U.S. patent application number 16/621538 was filed with the patent office on 2022-09-15 for heterocyclic compounds as mnk inhibitors.
The applicant listed for this patent is NANJING INNOCARE PHARMA TECH CO., LTD.. Invention is credited to Norman Xianglong Kong, Zhixiang Zheng, Chao Zhou.
Application Number | 20220289719 16/621538 |
Document ID | / |
Family ID | 1000006376958 |
Filed Date | 2022-09-15 |
United States Patent
Application |
20220289719 |
Kind Code |
A1 |
Kong; Norman Xianglong ; et
al. |
September 15, 2022 |
HETEROCYCLIC COMPOUNDS AS MNK INHIBITORS
Abstract
The present invention relates to a heterocyclic compound, a
pharmaceutical composition containing the same, a preparation
method thereof, and a use thereof as a mitogen-activated protein
kinase interacting kinase 1 and 2(MNK1/MNK2) inhibitor. The present
invention further relates to a method of treating or prevent
MNK-mediated diseases, such as cancer. The compound is a
heterocyclic compound as shown in Formula (I), or a
pharmaceutically acceptable salt, a prodrug, a solvate, a
polymorph, an isomer, or a stable isotopic derivative thereof.
##STR00001##
Inventors: |
Kong; Norman Xianglong;
(Nanjing, CN) ; Zhou; Chao; (Nanjing, CN) ;
Zheng; Zhixiang; (Nanjing, CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
NANJING INNOCARE PHARMA TECH CO., LTD. |
Nanjing, Jiangsu |
|
CN |
|
|
Family ID: |
1000006376958 |
Appl. No.: |
16/621538 |
Filed: |
June 8, 2018 |
PCT Filed: |
June 8, 2018 |
PCT NO: |
PCT/CN2018/090353 |
371 Date: |
December 11, 2019 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 487/04 20130101;
A61P 35/00 20180101; C07D 471/04 20130101; C07D 403/12 20130101;
C07D 403/14 20130101; C07D 401/14 20130101 |
International
Class: |
C07D 403/12 20060101
C07D403/12; C07D 401/14 20060101 C07D401/14; C07D 403/14 20060101
C07D403/14; C07D 471/04 20060101 C07D471/04; C07D 487/04 20060101
C07D487/04; A61P 35/00 20060101 A61P035/00 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 12, 2017 |
CN |
201710439265.5 |
Claims
1. A compound as shown in Formula (I), or an isomer, a prodrug, a
solvate, a stable isotopic derivative and a pharmaceutically
acceptable salt thereof: ##STR00121## Where R.sup.1, R.sup.2,
R.sup.3 are each independently selected from the group consisting
of hydrogen, halogen, cyano, C1-C8 alkyl, C3-C8 cyclyl, 3-8
membered heterocyclyl, aryl, heteroaryl, --C(O)R.sup.4, alkenyl,
alkynyl, --OR.sup.4, --NR.sup.5 R.sup.6, --OC(O)NR.sup.5R.sup.6,
--C(O)OR.sup.4, --C(O)NR.sup.5R.sup.6, --NR.sup.5C(O)R.sup.4,
--NR.sup.4C(O)NR.sup.5R.sup.6, --S(O).sub.mR.sup.4,
--NR.sup.5S(O).sub.mR.sup.4, --SR.sup.4,
--S(O).sub.mNR.sup.5R.sup.6, --NR.sup.4S(O).sub.mNR.sup.5R.sup.6,
where said alkyl, alkenyl, alkynyl, cyclyl, hetercyclyl, aryl,
heteroaryl are optionally substituted by one or more substituents
selected from halogen, cyano, C1-C8 alkyl, C3-C8 cyclyl,
3-8-membered heterocyclyl, --OR.sup.7, --OC(O)NR.sup.8R.sup.9,
--C(O)OR.sup.7, --C(O)NR.sup.8R.sup.9, --C(O)R.sup.7,
--NR.sup.8R.sup.9, --NRC(O)R.sup.7, --NR.sup.7C(O)NR.sup.8R.sup.9,
--S(O)mR.sup.7, --NR.sup.8S(O)mR.sup.7, --SR.sup.7,
--S(O)mNR.sup.8R.sup.9, --NR'S(O)mNR.sup.8R.sup.9; Where ring Ar
are each independently selected from substituted or unsubstituted
aryl or heteroaryl, when Ar is substituted, it could be substituted
by one or more substituents at any position, where said
substituents are each independently selected from the group
consisting of hydrogen, halogen, cyano, C1-C8 alkyl, C3-C8 cyclyl,
3-8 membered heterocyclyl, aryl, heteroaryl, --C(O)R.sup.4,
--C(O)OR.sup.4, alkenyl, alkynyl, OR.sup.4, --NR.sup.5R.sup.4,
--NR.sup.5C(O)R.sup.4, --NR.sup.4C(O)NR.sup.5R.sup.6,
--S(O)mR.sup.4, --NR.sup.5S(O)mR.sup.4, --SR.sup.4,
--S(O)mNR.sup.5R.sup.6, --NR.sup.4S(O)mNR.sup.5R.sup.6, where said
alkyl, alkenyl, alkynyl, cyclyl, hetercyclyl, aryl, heteroaryl are
optionally substituted by one or more substituents selected from
the group consisting of halogen, cyano, C1-C8 alkyl, C3-C8 cyclyl,
3-8 membered heterocyclyl, --OR.sup.7, --OC(O)NR.sup.8R.sup.9,
--C(O)OR.sup.7, --C(O)NR.sup.8R.sup.9, --C(O)R.sup.7,
--NR.sup.8R.sup.9, --NR.sup.8C(O)R.sup.7,
--NR.sup.7C(O)NR.sup.8R.sup.9, --S(O)mR.sup.1,
--NR.sup.8S(O)mR.sup.7, --SR.sup.7, --S(O)mNR.sup.8R.sup.9,
--NR.sup.7S(O)mNR.sup.8R.sup.9. R.sup.1 and R.sup.2 may form a 5-8
membered heterocyclyl together with the carbon atom to which they
are attached; R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.1, R.sup.9
are each independently selected from the group consisting of
hydrogen, C1-C8 alkyl, heteroalkyl, C3-C8 cyclyl, 3-8 membered
monocyclic heterocyclyl, monocyclic heteroaryl or monocyclic aryl,
alkenyl, alkynyl, where said R.sup.5 and R.sup.6, R.sup.8 and
R.sup.9 may form a 3-7 membered heterocyclyl together with the
carbon atom to which they are attached; and m is 1 or 2.
2. The compound according to claim 1, an isomer, a prodrug, a
solvate, a stable isotopic derivative thereof or a pharmaceutically
acceptable salt thereof, characterized in that the compound is of
the following formula (II) a-g: ##STR00122## where: R.sup.1,
R.sup.2, R.sup.3, R.sup.10, R.sup.11, R.sup.13, R.sup.14, R.sup.15
are each independently selected from the group consisting of
hydrogen, halogen, cyano, C1-C8 alkyl, C3-C8 cyclyl, 3-8 membered
heterocyclyl, aryl, heteroaryl, aldehyde group, --C(O)R.sup.4,
carboxyl, alkenyl, alkynyl, --OR.sup.4, --NR.sup.5R.sup.6,
--NR.sup.5C(O)R.sup.4, --NR.sup.4C(O)NR.sup.5R.sup.6,
--S(O)mR.sup.4, --NR.sup.5S(O)mR.sup.4, --SR.sup.4,
--S(O)mNR.sup.5R.sup.6, --NR.sup.4S(O)mNR.sup.5R.sup.6, where said
alkyl, cyclyl, heterocyclyl, aryl, or heteroaryl are optionally
substituted by one or more substituents selected from the group
consisting of halogen, cyano, C1-C8 alkyl, C3-C8 cyclyl, 3-8
membered heterocyclyl, --OR.sup.7, --OC(O)NR.sup.8R.sup.9,
--C(O)OR.sup.7, --C(O)NR.sup.8R.sup.9, --C(O)R.sup.7,
--NR.sup.8R.sup.9, --NR.sup.7C(O)R.sup.7,
--NR.sup.7C(O)NR.sup.8R.sup.9, --S(O)mR.sup.7, --NR'S(O)mR.sup.7,
--SR.sup.7, --S(O)mNR.sup.8R.sup.9, --NR.sup.7S(O)mNR.sup.8R.sup.9;
R.sup.2 was selected from the group consisting of hydrogen, C1-C8
alkyl, C3-C8 cyclyl, 3-8 membered monocyclic heterocyclyl,
monocyclic heteroaryl or monocyclic aryl, where said alkyl, cyclyl,
heterocyclyl, aryl or heteroaryl are optionally substituted by one
or more substituents selected from the group consisting of halogen,
cyano, C1-C8 alkyl, C3-C8 cyclyl, 3-8 membered heterocyclyl,
--OR.sup.7, --OC(O)NR.sup.8R.sup.9, --C(O)OR.sup.7,
--C(O)NR.sup.8R.sup.9, --C(O)R.sup.7, --NR.sup.8R.sup.9,
--NR.sup.7C(O)R.sup.1, --NR.sup.7C(O)NR.sup.8R.sup.9,
--S(O)mR.sup.7, --NR'S(O)mR.sup.7, --SR.sup.7,
--S(O)mNR.sup.8R.sup.9, --NR'S(O)mNR.sup.8R.sup.9; R.sup.16,
R.sup.7 are each independently selected from the group consisting
of hydrogen, C1-C8 alkyl, C3-C8 cyclyl, 3-8 membered monocyclic
heterocyclyl, monocyclic heteroaryl or monocyclic aryl, where said
alkyl, cyclyl, heterocyclyl, aryl or heteroaryl are optionally
substituted by one or more substituents selected from the group of
halogen, cyano, C1-C8 alkyl, C3-C8 cyclyl, 3-8 membered
heterocyclyl, --OR.sup.7, --OC(O)NR.sup.8R.sup.9, --C(O)OR.sup.7,
--C(O)NR.sup.8R.sup.9, --C(O)R.sup.7, --NR.sup.7R.sup.9,
--NR.sup.8C(O)R.sup.7, --NR.sup.7C(O)NR.sup.8R.sup.9,
--S(O)mR.sup.7, NR.sup.8S(O)mR.sup.7, --SR.sup.7,
--S(O)mNR.sup.8R.sup.9, --NR'S(O)mNR.sup.8R.sup.9. R.sup.1 and
R.sup.2 may form a 5-8 membered hetero-cyclyl with the atom they
are attached; R.sup.10 and R.sup.11 may form a 5-8 membered
heterocyclyl with the atom they are attached; R.sup.16 and R.sup.17
may form a 3-8 membered heterocyclyl with the atom they are
attached; the definition of R.sup.4-9 are described as above; and m
is 1 or 2.
3. The compound according to claim 1, an isomer, a prodrug, a
solvate, a stable isotopic derivative thereof or a pharmaceutically
acceptable salt thereof, characterized in that the compound is of
the following Formula (III) a-e ##STR00123## where: R.sup.2 is
selected from the group consisting of hydrogen, fluoro, cyano,
C1-C3 alkyl, C5-C6 cyclyl, 5-6 membered heterocyclyl, aryl,
heteroaryl, --C(O)OR.sup.4, --C(O)NR.sup.5R.sup.6, carboxyl,
--OR.sup.4, --NR.sup.5R.sup.6, where said cyclyl, heterocyclyl are
optionally substituted by one or more substituents selected from
the group consisting of --C(O)OR.sup.7, --C(O)NR.sup.8R.sup.9;
R.sup.12 is selected from hydrogen, alkoxycarbonyl, alkylcarbonyl,
cycloalkylcarbonyl; R.sup.18 is selected from hydrogen, C1-C5
alkyl, C3-C6 cycloalkyl, aryl, 5-6 membered heteroaryl,
alkylcarbonyl, cycloalkylcarbonyl, arylcarbonyl,
heteroarylcarbonyl, alkylsulfonyl, cycloalkylsulfonyl; R.sup.19 is
selected from C1-C4 alkyl; R.sup.20 is selected from C1-C5 alkyl,
C1-C5 oxaalkyl, --CH.sub.2CH.sub.2NR.sup.5R.sup.6, phenyl; R.sup.2
and R.sup.18 may form a 5-8 membered ring with the carbon and
nitrogen atoms they are attached; R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.1, R.sup.9 are each independently selected from the
group consisting of hydrogen, C1-C5 alkyl, C3-C8 cyclyl, 3-8
membered monocyclic heterocyclyl, monocyclic heteroaryl or
monocyclic aryl, alkenyl, alkynyl, where said R.sup.5 and R.sup.6,
R.sup.8 and R.sup.9 may form a 3-7 membered heterocyclyl with the
nitrogen they are attached.
4. The compound according to claim 1, an isomer, a prodrug, a
solvate, a stable isotopic derivative thereof or a pharmaceutically
acceptable salt thereof, characterized in that the compound is of
Formula (III) f-g: ##STR00124## where, R.sup.2 is selected from the
group consisting of hydrogen, fluoro, cyano, C1-C3 alkyl, C5-C6
cyclyl, 5-6 membered heterocyclyl, aryl, heteroaryl,
--C(O)OR.sup.4, --C(O)NR.sup.5R.sup.6, carboxyl, --OR.sup.4,
--NR.sup.5R.sup.6, where said cyclyl, heterocyclyl are optionally
substituted by substituents selected from --C(O)OR.sup.7,
--C(O)NR.sup.8R.sup.9; R.sup.12 is selected from C1-C6 alkyl, C5-C6
cyclyl; R.sup.16, R.sup.17 are each independently selected from
C1-C5 alkyl; R.sup.18 is selected from hydrogen, C1-C5 alkyl, C3-C6
cycloalkyl, aryl, 5-6 membered heteroaryl, alkylcarbonyl,
cycloalylcarbonyl, arylcarbonyl, heteroarylcarbonyl; R.sup.19 is
selected from C1-C4 alkyl; R.sup.20 is selected from C1-C5 alkyl,
C1-C5 oxaalkyl, --CH.sub.2CH.sub.2NR.sup.5R.sup.6; R.sup.2 and
R.sup.18 may form a 5-8 membered heterocyclyl with the atom they
are attached; R.sup.16 and R.sup.17 may form a 4-6 membered ring
with the atom they are attached; R.sup.4, R.sup.5, R.sup.6,
R.sup.7, R.sup.1, R.sup.9 are each independently selected from the
group consisting of hydrogen, C1-C5 alkyl, C3-C8 cyclyl, 3-8
membered monocyclic heterocyclyl, monocyclic heteroaryl or
monocyclic aryl, alkenyl, alkynyl, where said R.sup.5 and R.sup.6,
R.sup.8 and R.sup.9 may form a 3-7 membered heterocyclyl with the
nitrogen atoms they are attached.
5. The compound according to claim 1, an isomer, a prodrug, a
solvate, a stable isotopic derivative thereof or a pharmaceutically
acceptable salt thereof, which is selected from: ##STR00125##
##STR00126## ##STR00127## ##STR00128## ##STR00129## ##STR00130##
##STR00131## ##STR00132## ##STR00133## ##STR00134## ##STR00135##
##STR00136## ##STR00137##
6. A pharmaceutical composition, comprising a compound according to
claim 1 or an isomer, a prodrug, a stable isotopic derivative or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier, diluent and excipient.
7. (canceled)
8. A method for treating or preventing MNK-mediated diseases,
comprising administering to a patient in need thereof a
therapeutically effective amount of a compound according to claim 1
or an isomer thereof, a prodrug, a solvate, a stable isotope
derivative or a pharmaceutically acceptable salt thereof.
9. (canceled)
10. The method of claim 8, wherein the MNK-mediated diseases are
selected from malignant hematological diseases, lung cancer, breast
cancer, ovarian cancer, prostate cancer, pancreatic cancer, and
glioma.
Description
TECHNICAL FIELD
[0001] The present invention relates to a class of heterocyclic
compounds, the preparation method thereof, a pharmaceutical
composition containing the same, and a use thereof as a MAP
kinase-interacting kinase (MNK) inhibitor. The compounds according
to the present invention can be used to treat or prevent related
diseases mediated by MNK, such as cancer.
BACKGROUND ART
[0002] The mitogen-activated protein kinase-interacting kinase
(MNK) belongs to the serine/threonine protein kinase and was first
discovered in 1997 as an extracellular regulatory protein kinase
(ERK) substrate or binding factor (Waskiewicz A. et al. EMBO J.,
1997, 16(8), 1909-1920 & 1921-1933).
[0003] Human MNK protein is encoded by two groups of genes, MKNK1
and MKNK2. Each group of genes are translated into two subtypes by
selective splicing, namely MNK1a, MNK1b and MNK2a, MNK2b. From the
protein sequence analysis, each of these four subtypes contains a
nuclear localization signal (NLS) at the N-terminus, and a sequence
that binds to eIF4G, which allows MNK kinase to enter the nucleus
and function. A kinase domain with high sequence homology therein
is responsible for the catalytic function of the kinase. This
kinase domain, which belongs to the Ca+/calmodulin-regulated
protein kinase (CaMK) family, is not affected by selective splicing
and has a high degree of conservation between the four subtypes.
The main structural difference is at the C-terminus, MNK1a and
MNK2a contain a MAPK domain at the C-terminus, which is responsible
for the activation of upstream ERK and p38. Whereas this domain is
deleted in the other two subtypes which cannot be phosphorylated
and activated by the upstream kinase, therefore, they have
different basal activities. MNK1a has a nuclear export signal (NES)
at the C-terminus, which allows the MNK1a subtype to be more widely
distributed in the cytoplasm, while the other three subtypes are
mostly present in nucleus (Diab S. et. al. Chem. Biol. 2014, 21(4),
441-452). eIF4E is the protein which has been studied earliest and
most comprehensively in the substrates discovered so far. Through
the eIF4E binding domain of N-terminal, MNK1/2 can bind to eIF4E,
and then phosphorylates its serine at position 209, and regulates
the translation process of related proteins. These proteins play an
important role in the mechanisms of tumor cell pro-survival,
anti-apoptosis, metastasis and drug resistance. MNK overactivation
marked by up-regulation of p-eIF4E level can be detected in
prostate cancer, breast cancer, pancreatic cancer, lung cancer,
glioma, leukemia, etc. (Lim S. et. al. Proc. Natl. Acad. Sci. USA,
2013, 110(20), 2298-2307; Grzmil. M. et al. J. Clin. Invest., 2014,
124(2), 742-754; Yoshizawa A. et. al. Clin. Cancer Res. 2010, 16
(1), 204-248; Adesso L. et. al. Oncogene, 2012, 32(23), 2848-2857
et al.).
[0004] MNK is a kinase which acts downstream of the MAPK pathway,
and its pro-survival effect is mainly dependent on the promotion of
translation process of tumor-associated proteins. Studies have
confirmed that MNK can promote the translation of related mRNA,
promote tumor angiogenesis and cell proliferation, and inhibit
apoptosis. Recent studies have shown that MNK maintains tumor cell
survival in diffuse large B-cell lymphoma (DLBCL), and inhibition
of MNK can not only block eIF4E1 phosphorylation, but also enhance
eIF4E3 expression (Landon A. et. al. Nat. Commun. 2014, 5,
5413.).
[0005] Studies of molecular mechanisms have indicated that
MNK-mediated up-regulation of eIF4E phosphorylation promotes
translation of Snail and MMP-3 proteins, induces
epithelial-mesenchymal transition (EMT), and thereby promotes tumor
metastasis. The inhibition of MNK and eIF4E phosphorylation
mediated by MNK is expected to be an effective solution for tumor
metastasis (Robichaud N. et. al. Oncogene, 2014, 34(16),
2032-2042).
[0006] Studies have confirmed that MNK kinase plays a role in
multiple drug-induced compensatory pathways, which ultimately leads
to drug resistance. The resistance to mTOR inhibitor rapamycin and
its analogues is related to MNK. The combination of MNK inhibitor
and rapamycin can overcome this drug resistance pathway and produce
a synergistic effect, effectively block the translation level of
related proteins and inhibit the proliferation of tumor cells to
give a better anti-tumor effect. Studies have indicated that the
resistances to other chemotherapeutic drugs, such as imatinib,
cytarabine, gemcitabine, etc. are all associated with MNK and eIF4E
phosphorylation levels. The combination of these drugs with MNK
inhibitors may effectively reverse the drug resistance.
[0007] Therefore, the combination of MNK inhibitors with some
clinically standard therapeutics is an effective treatment strategy
(Adesso L. et. al. Oncogene, 2012, 32(23), 2848-2857; Lim S. et al.
Proc. Natl. Acad. Sci. USA, 2013, 110(20), 2298-2307; Altman J. K.
et. al. Mol. Pharmacol. 2010, 78(4), 778-784).
[0008] Therefore, this invention provides novel MNK inhibitor
compounds.
SUMMARY OF THE INVENTION
[0009] The purpose of the present invention is to provide a
compound as shown in Formula (I), or an isomer, a prodrug, a
solvate, a stable isotopic derivative and a pharmaceutically
acceptable salt thereof:
##STR00002##
Where
[0010] R.sup.1, R.sup.2, R.sup.3 are each independently selected
from the group consisting of hydrogen, halogen, cyano, C1-C8 alkyl,
C3-C8 cyclyl, 3-8 membered heterocyclyl, aryl, heteroaryl, aldehyde
group, --C(O)R.sup.4, carboxyl, alkenyl, alkynyl, --OR.sup.4,
--NR.sup.5R.sup.6, --OC(O)NR.sup.5R.sup.6, --C(O)OR.sup.4,
--C(O)NR.sup.5R.sup.6, --NR.sup.5C(O)R.sup.4,
--NR.sup.4C(O)NR.sup.5R.sup.6, --S(O).sub.mR.sup.4,
--NR.sup.5S(O).sub.mR.sup.4, --SR.sup.4,
--S(O).sub.mNR.sup.5R.sup.6, --NR.sup.4S(O).sub.mNR.sup.5R.sup.6,
where said alkyl, cyclyl, hetercyclyl, aryl or heteroaryl are
optionally substituted by one or more substituents selected from
halogen, cyano, C1-C8 alkyl, C3-C8 cyclyl, 3-8-membered
heterocyclyl, --OR.sup.7, --OC(O)NR.sup.8R.sup.9, --C(O)OR.sup.7,
--C(O)NR.sup.8R.sup.9, --C(O)R.sup.7, --NR.sup.8R.sup.9,
--NR.sup.8C(O)R.sup.7, --NR.sup.7C(O)NR.sup.8R.sup.9,
--S(O).sub.mR.sup.7, --NR.sup.8S(O).sub.mR.sup.7, --SR.sup.7,
--S(O).sub.mNR.sup.8R.sup.9,
--NR.sup.7S(O).sub.mNR.sup.8R.sup.9;
[0011] Wherein ring Ar are each independently selected from
substituted or unsubstituted aryl or heteroaryl, when Ar is
substituted, it could be substituted by one or more substituents at
any position, said substituents are each independently selected
from the group consisting of hydrogen, halogen, cyano, C1-C8 alkyl,
C3-C8 cyclyl, 3-8 membered heterocyclyl, aryl, heteroaryl, aldehyde
group, --C(O)R.sup.4, carboxyl, alkenyl, alkynyl, OR.sup.4,
--NR.sup.5R.sup.6'--NR.sup.5C(O)R.sup.4,
--NR.sup.4C(O)NR.sup.5R.sup.6, --S(O).sub.mR.sup.4,
--NR.sup.5S(O).sub.mR.sup.4, --SR.sup.4,
--S(O).sub.mNR.sup.5R.sup.6, --NR.sup.4S(O).sub.mNR.sup.5R.sup.6,
wherein said alkyl, cyclyl, hetercyclyl, aryl, heteroaryl are
optionally substituted by one or more substituents selected from
the group consisting of halogen, cyano, C1-C8 alkyl, C3-C8 cyclyl,
3-8 membered heterocyclyl, --OR.sup.7, --OC(O)NR.sup.8R.sup.9,
--C(O)OR.sup.7, --C(O)NR.sup.8R.sup.9, --C(O)R.sup.7,
--NR.sup.8R.sup.9, --NR.sub.8C(O)R.sup.8,
--NR.sup.4C(O)NR.sup.8R.sup.9, --S(O).sub.mR.sup.7,
--NR.sup.8S(O).sub.mR.sup.7, --SR.sup.7,
--S(O).sub.mNR.sup.8R.sup.9,
--NR.sup.7S(O).sub.mNR.sup.8R.sup.9.
[0012] R.sup.1 and R.sup.2 may form a 5-8 membered heterocyclyl
together with the carbon atom to which they are attached;
[0013] R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9 are
each independently selected from the group consisting of hydrogen,
C1-C8 alkyl, heteroalkyl, C3-C8 cyclyl, 3-8 membered monocyclic
heterocyclyl, monocyclic heteroaryl or monocyclic aryl, alkenyl,
alkynyl, wherein said R.sup.5 and R.sup.6, R.sup.8 and R.sup.9 may
form a 3-7 membered heterocyclyl; and m is 1 or 2.
[0014] In one embodiment of the present invention, a compound as
shown in general formula (I), an isomer, a prodrug, a solvate, a
stable isotopic derivative thereof or a pharmaceutically acceptable
salt thereof is provided, characterized in that the compound s have
structures of the following Formula (II);
##STR00003##
[0015] wherein:
[0016] R.sup.1, R.sup.2, R.sup.3, R.sup.10, R.sup.11, R.sup.13,
R.sup.14, R.sup.15 are each independently selected from the group
consisting of hydrogen, halogen, cyano, C1-C8 alkyl, C3-C8 cyclyl,
3-8 membered heterocyclyl, aryl, heteroaryl, aldehyde group,
--C(O)R.sup.4, carboxyl, alkenyl, alkynyl, --OR 4,
--NR.sup.5R.sup.6, --NR.sup.5C(O)R.sup.4,
--NR.sup.4C(O)NR.sup.5R.sup.6, --S(O).sub.mR.sup.4,
--NR.sup.5S(O).sub.m, R.sup.4, --SR.sup.4, --S(O).sub.m,
NR.sup.5R.sup.6, --NR.sup.4S(O).sub.m, NR.sup.5R.sup.6, wherein
said alkyl, cyclyl, heterocyclyl, aryl, or heteroaryl are
optionally substituted by one or more substituents selected from
the group consisting of halogen, cyano, C1-C8 alkyl, C3-C8 cyclyl,
3-8 membered heterocyclyl, --OR.sup.1, --OC(O)NR.sup.8R.sup.9,
--C(O)OR.sup.7, --C(O)NR.sup.8R.sup.9, --C(O)R.sup.7,
--NR.sup.8R.sup.9, --NR.sup.8C(O)R.sup.7,
--NR.sup.7C(O)NR.sup.8R.sup.9, --S(O).sub.mR.sup.7,
--NR.sup.8S(O).sub.mR.sup.7, --SR.sup.7,
--S(O).sub.mNR.sup.8R.sup.9,
--NR.sup.7S(O).sub.mNR.sup.8R.sup.9;
[0017] R.sup.2 was selected from the group consisting of hydrogen,
C1-C8 alkyl, C3-C8 cyclyl, 3-8 membered monocyclic heterocyclyl,
monocyclic heteroaryl or monocyclic aryl, wherein said alkyl,
cyclyl, heterocyclyl, aryl or heteroaryl are optionally substituted
by one or more substituents selected from the group consisting of
halogen, cyano, C1-C8 alkyl, C3-C8 cyclyl, 3-8 membered
heterocyclyl, --OR.sup.7, --OC(O)NR.sup.8R.sup.9, --C(O)OR.sup.7,
--C(O)NR.sup.8R.sup.9, --C(O)R.sup.7, --NR.sup.8R.sup.9,
--NR.sup.8C(O)R.sup.7, --NR.sup.7C(O)NR.sup.8R.sup.9,
--S(O).sub.mR.sup.7, --NR.sup.8S(O).sub.mR.sup.7, --SR.sup.7,
--S(O).sub.mNR.sup.8R.sup.9,
--NR.sup.7S(O).sub.mNR.sup.8R.sup.9;
[0018] R.sup.16, R.sup.17 are each independently selected from the
group consisting of hydrogen, C1-C8 alkyl, C3-C8 cyclyl, 3-8
membered monocyclic heterocyclyl, monocyclic heteroaryl or
monocyclic aryl, wherein said alkyl, cyclyl, heterocyclyl, aryl or
heteroaryl are optionally substituted by one or more substituents
selected from the group consisting of halogen, cyano, C1-C8 alkyl,
C3-C8 cyclyl, 3-8 membered heterocyclyl, --OR.sup.7,
--OC(O)NR.sup.8R.sup.9, --C(O)OR.sup.7, --C(O)NR.sup.8R.sup.9,
--C(O)R.sup.7, --NR.sup.8R.sup.9, --NR.sup.8C(O)R.sup.7,
--NR.sup.7C(O)NR.sup.8R.sup.9, --S(O).sub.mR.sup.7,
--NR.sup.8S(O).sub.mR.sup.7, --SR.sup.7,
--S(O).sub.mNR.sup.8R.sup.9,
--NR.sup.7S(O).sub.mNR.sup.8R.sup.9.
[0019] R.sup.1 and R.sup.2 may form a 5-8 membered heterocyclyl
with the atom they are attached;
[0020] R.sup.10 and R.sup.11 may form a 5-8 membered heterocyclyl
with the atom they are attached;
[0021] R.sup.16 and R.sup.17 may form a 5-8 membered heterocyclyl
with the atom they are attached;
[0022] The definition of R.sup.4-9 are as described above;
[0023] And m is 1 or 2.
[0024] In another embodiment of the present invention, a compound
as shown in general formula (I) or (II), an isomer, a prodrug, a
solvate, a stable isotopic derivative thereof or a pharmaceutically
acceptable salt thereof is provided, characterized in that the
compound has structure as shown in the following Formula (III)
a-e
##STR00004##
[0025] In Formula (III) a-e:
[0026] R.sup.2 is selected from the group consisting of hydrogen,
fluoro, cyano, C1-C3 alkyl, C5-C6 cyclyl, 5-6 membered
heterocyclyl, aryl, heteroaryl, --C(O)OR.sup.4,
--C(O)NR.sup.5R.sup.6, carboxyl, --OR.sup.4, --NR.sup.5R.sup.6,
wherein said cyclyl, heterocyclyl are optionally substituted by one
or more substituents selected from the group consisting of
--C(O)OR.sup.7, --C(O)NR.sup.8R.sup.9; R.sup.12 is selected from
hydrogen,
[0027] alkoxycarbonyl, alkylcarbonyl, cycloalkylcarbonyl; R.sup.18
is selected from
[0028] hydrogen, C1-C5 alkyl, C3-C6 cycloalkyl, aryl, 5-6 membered
heteroaryl, alkylcarbonyl, cycloalkylcarbonyl, arylcarbonyl,
heteroarylcarbonyl, alkylsulfonyl, cycloalkylsulfonyl; R.sup.19 is
selected from C1-C4 alkyl; R.sup.20 is selected from C1-C5
[0029] alkyl, C1-C5 oxaalkyl, --CH.sub.2CH.sub.2NR.sup.5R.sup.6,
phenyl;
[0030] R.sup.2 and R.sup.18 may form a nitrogen atom-containing 5-8
membered ring with the carbon and nitrogen atoms they are
attached;
[0031] R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9 are
each independently selected from the group consisting of hydrogen,
C1-C5 alkyl, C3-C8 cyclyl, 3-8 membered monocyclic heterocyclyl,
monocyclic heteroaryl or monocyclic aryl, alkenyl, alkynyl, where
said R.sup.5 and R.sup.6, R.sup.8 and R.sup.9 may form a 3-7
membered heterocyclyl with the nitrogen they are attached.
[0032] In another embodiment of the present invention, a compound
as shown in general formula (I) or (II), an isomer, a prodrug, a
solvate, a stable isotopic derivative thereof or a pharmaceutically
acceptable salt thereof is provided, characterized in that the
compound has structure as shown in the following Formula (III)
f-g
##STR00005##
[0033] In formula IIIf-g
[0034] R.sup.2 is selected from the group consisting of hydrogen,
fluoro, cyano, C1-C3 alkyl, C5-C6 cyclyl, 5-6 membered
heterocyclyl, aryl, heteroaryl, --C(O)OR.sup.4,
--C(O)NR.sup.5R.sup.6, carboxyl, --OR.sup.4, --NR.sup.5R.sup.6,
where said cyclyl, heterocyclyl are optionally substituted by one
substituent selected from --C(O)OR.sup.7,
--C(O)NR.sup.8R.sup.9;
[0035] R.sup.12 is selected from C1-C6 alkyl, C5-C6 cyclyl.
[0036] R.sup.16, R.sup.17 are each independently selected from
C1-C5 alkyl;
[0037] R.sup.18 is selected from hydrogen, C1-C5 alkyl, C3-C6
cycloalkyl, aryl, 5-6 membered heteroaryl, alkylcarbonyl,
cycloalkylcarbonyl, arylcarbonyl, heteroarylcarbonyl;
[0038] R.sup.19 is selected from C1-C4 alkyl;
[0039] R.sup.20 is selected from C1-C5 alkyl, C1-C5 oxaalkyl,
--CH.sub.2CH.sub.2NR.sup.5R.sup.6;
[0040] R.sup.2 and R.sup.18 may form a 5-8 membered heterocyclyl
with the atom they are attached;
[0041] R.sup.16 and R.sup.17 may form a 4-6 membered ring with the
atom they are attached;
[0042] R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9 are
each independently selected from the group consisting of hydrogen,
C1-C5 alkyl, C3-C8 cyclyl, 3-8 membered monocyclic heterocyclyl,
monocyclic heteroaryl or monocyclic aryl, alkenyl, alkynyl, where
said R.sup.5 and R.sup.6, R.sup.8 and R.sup.9 may form a 3-7
membered heterocyclyl with the nitrogen atom they are attached.
[0043] The preferred compounds according to the present invention
include, but not limited to
##STR00006## ##STR00007## ##STR00008## ##STR00009## ##STR00010##
##STR00011## ##STR00012## ##STR00013## ##STR00014## ##STR00015##
##STR00016## ##STR00017## ##STR00018## ##STR00019## ##STR00020##
##STR00021## ##STR00022##
and an isomer, a prodrug, a solvate, a stable isotopic derivative
and a pharmaceutically acceptable salt thereof.
[0044] The present invention further relates to a pharmaceutical
composition comprising a compound of the present invention or an
isomer, a prodrug, a stable isotopic derivative or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier, diluent and excipient.
[0045] In another aspect, the present invention relates to use of
the compound of formula (I) or an isomer, a prodrug, a stable
isotopic derivative or a pharmaceutically acceptable salt thereof,
or the pharmaceutical composition in the manufacture of a
medicament, wherein the medicament is used in the treatment or
prevention of MNK-mediated diseases, such as tumors, especially
malignant hematological diseases, lung cancer, breast cancer,
ovarian cancer, prostate cancer, pancreatic cancer, glioma.
[0046] In another aspect, the present invention relates to use of
the compound of formula (I) or an isomer, a prodrug, a stable
isotopic derivative or a pharmaceutically acceptable salt thereof,
or the pharmaceutical composition in the manufacture of a
medicament for the treatment or prevention of diseases such as
cancers and inflammation.
[0047] According to the present invention, the medicament can be in
any dosage form, including but not limited to tablets, capsules,
solution, lyophilized preparation and injection.
[0048] The pharmaceutical preparation of the present invention may
be administered in the form of a dosage unit containing a
predetermined amount of an active ingredient per dosage unit. Such
a unit may contain, for example, 0.5 mg to 1 g, preferably 1 mg to
700 mg, particularly preferably 5 mg to 300 mg of a compound of the
present invention, or a drug depending on the condition to be
treated, the method of administration and the age, weight and
condition of the patient, or the pharmaceutical preparation may be
administered in the form of a dosage unit containing a
predetermined amount of the active ingredient per dosage unit.
Preferred dosage unit formulations are those containing the daily
or divided dose or active fraction thereof as indicated above. In
addition, this type of pharmaceutical preparation can be prepared
using methods known in the pharmaceutical art.
[0049] The pharmaceutical formulations of the present invention may
be suitable for administration by any desired suitable method, such
as oral (including oral or sublingual), rectal, nasal, topical
(including oral, sublingual or transdermal), vaginal or parenteral
(Including subcutaneous, intramuscular, intravenous or intradermal)
administration. Such formulations can be prepared using all methods
known in the pharmaceutical art, for example, by combining the
active ingredient with one or more excipients or one or more
adjuvants.
[0050] The present invention also relates to a method for treating
or preventing MNK-mediated diseases (such as tumors, especially
malignant hematological diseases, lung cancer, breast cancer,
ovarian cancer, prostate cancer, pancreatic cancer, glioma),
comprising administering to a patient in need thereof
therapeutically effective amount of a compound of the present
invention or an isomer, a prodrug, a solvate, a stable isotope
derivative or a pharmaceutically acceptable salt thereof, or a
pharmaceutical composition of the present invention.
[0051] Another aspect of the present invention relates to a
compound of the general formula (I) or an isomer, a prodrug, a
solvate, a stable isotope derivative or a pharmaceutically
acceptable salt thereof, or a pharmaceutical composition, for use
in treating or preventing MNK-mediated diseases, such as tumors,
especially malignant hematological diseases, lung cancer, breast
cancer, ovarian cancer, prostate cancer, pancreatic cancer,
glioma.
[0052] Another aspect of the present invention relates to a
compound of the general formula (I) or tautomers, mesomers,
racemates, enantiomers, diastereomers, mixtures thereof, and
pharmaceutically acceptable salts thereof for use in treating
and/or preventing diseases such as cancer and so on.
Preparation Schemes
[0053] The present invention further provides methods for preparing
the Compounds.
##STR00023##
[0054] The definitions of R.sup.1, R.sup.2, R.sup.12, R.sup.18,
R.sup.19 are the same as described before.
[0055] Step 1:
[0056] X.sup.1, X.sup.2 are leaving groups such as halogen (Cl, Br,
I), OTf, OTs, OMs etc.
[0057] The Buchwald reaction is carried out in 1,4-dioxane,
N,N-dimethylacetamide or the like. Cesium carbonate or sodium
t-butoxide is added as a base. The catalyst used is
tris(dibenzylideneacetone) di-palladium or palladium acetate. The
ligand used is 4,5-bisdiphenylphosphino-9,9-dimethyloxaxene or
2-(dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphen-
yl, etc. The reaction is carried out in microwave or oil bath at
110-150.degree. C. The reaction gives compound (II);
[0058] Step 2:
[0059] X.sup.2 is a leaving group such as halogen (Cl, Br, I), OTf,
OTs, OMs etc. PG is an amino protecting group such as
t-butoxycarbonyl, trimethylsilylethoxymethyl, Y is CH or N. Y and
PG are unchanged during the reaction. Buchwald reaction is carried
out in 1,4-dioxane, N, N-dimethylacetamide, etc. Cesium carbonate
or sodium t-butoxide is added as the base. The catalyst used is
tris(dibenzylideneacetone)dipalladium or palladium acetate. The
ligand used is 4,5-bis-diphenylphosphino-9,9-dimethyloxaxene or
2-(dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphen-
yl, etc. The reaction is carried out in microwave oven or oil bath
at 110 to 150.degree. C. The reaction gives compound (Ill);
[0060] Step 3:
[0061] Y is unchanged during the reaction. Strong acid such as
trifluoroacetic acid/dichloromethane or hydrochloric acid/1,
4-dioxane is employed as the de-protection reagent. The reaction is
carried out under room temperature or by heating, PG protection
group is removed to give compound (IV).
[0062] Step 4:
[0063] Acetyl chloride or halide is used in the reaction. Y is
unchanged during the reaction. The solvent used is selected from
tetrahydrofuran, N,N-dimethylformamide. Base such as cesium
carbonate or sodium hydride is added. The reaction is carried out
at room temperature or by heating to give compound Scheme 2
##STR00024##
[0064] The definitions of R.sup.1, R.sup.2, R.sup.12, R.sup.18,
R.sup.19 are the same as described before.
[0065] Step 1:
[0066] X.sup.1 is a leaving group such as halogen (Cl, Br or I),
OTf, OTs, OMs etc. Buchwald reaction is carried out in 1,4-dioxane
or N, N-dimethylacetamide. Base such as cesium carbonate or t-BuONa
is added. The catalyst used is
tris(dibenzylideneacetone)dipalladium or palladium acetate. The
ligand used is 4,5-bisdiphenylphosphino-9,9-dimethyloxaxene or
2-(dicyclohexylphosphine)-3,6-dimethoxy-2',4',
6'-triisopropyl-1,1'-biphenyl etc.. The reaction takes place at
110.about.150.degree. C. in an oil bath or under microwave
conditions to give Compound(VII).
[0067] Step 2:
[0068] X.sup.2 is a leaving group such as halogen (Cl, Br, I), OTf,
OTs, OMs etc. PG is t-butoxycarbonyl, trimethylsilylethoxymethyl,
etc. Y is selected from N or CH. Y and PG are unchanged during the
reaction. Buchwald reaction is carried out in 1,4-dioxane or
N,N-dimethylacetamide. Cesium carbonate or sodium t-butoxide is
added at the same time as a base. The catalyst used was
tris(dibenzylideneacetone)dipalladium or palladium acetate. The
ligand used is 4,5-bisdiphenylphosphino-9,9-dimethyloxaxene or
2-(dicyclohexyl)
phosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl, etc.
The reaction takes place at 110.about.150.degree. C. in an oil bath
or under microwave conditions to give Compound(III).
[0069] Step 3:
[0070] Y is unchanged during the reaction. Strong acid such as
trifluoroacetic acid/dichloromethane or hydrochloric acid/1,
4-dioxane is employed as the de-protection reagent. The reaction is
carried out at room temperature or by heating.
[0071] The protection group is removed to give compound (IV).
[0072] Step 4:
[0073] Acetyl chloride or halide is used in the reaction. Y is
unchanged during the reaction. The solvent of the reaction is
selected from tetrahydrofuran or N,N-dimethylformamide. Base such
as cesium carbonate or sodium hydride is added. The reaction is
carried out at room temperature or by heating to give compound
(V).
##STR00025##
[0074] The definitions of R.sup.12, R.sup.19, R.sup.20 are the same
as described before.
[0075] Step 1:
[0076] X.sup.1 is a leaving group such as halogen (Cl, Br or I),
OTf, OTs or OMs, etc. L is selected from O or NH. The solvent of
the reaction is N, N-dimethylformamide.
[0077] Base such as lithium hydroxide is added. The reaction takes
place at 60.degree. C. to give compound (IX).
[0078] Step 2:
[0079] X.sup.2 is a leaving group such as halogen (Cl, Br, I), OTf,
OTs or OMs etc. L is selected from O or NH. PG is t-butoxycarbonyl,
trimethylsilylethoxymethyl, etc. L and PG are unchanged during the
reaction. Buchwald reaction is carried out in such as 1,4-dioxane
or N,N-dimethylacetamide. At the same time, cesium carbonate or
sodium t-butoxide is added as a base, and the catalyst used is
tris(dibenzylideneacetone)dipalladium or palladium acetate. The
ligand used is 4,5-bisdiphenylphosphine-9,9-dimethyl xanthene or
2-(dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphen-
yl, etc. The reaction takes place at 110.about.150.degree. C. in an
oil bath or under microwave conditions to give compound(X).
[0080] Step 3:
[0081] L is selected from O or NH. Strong acid such as
trifluoroacetic acid/dichloromethane or hydrochloric acid/1,
4-dioxane is employed as the de-protection reagent. L is unchanged
during the reaction. The reaction is carried out at room
temperature or by heating. Protection group is removed to give
compound (XI).
[0082] Step 4:
[0083] L is selected from O or NH. Corresponding acyl chloride or
acyl halide is used in the reaction. L is unchanged during the
reaction. The solvent of the reaction is selected from
tetrahydrofuran, N,N-dimethylformamide. Base such as cesium
carbonate or sodium hydride is added, the reaction is carried out
at room temperature or by heating to give compound (XII).
##STR00026##
[0084] The definitions of R.sup.2, R.sup.12, R.sup.16, R.sup.17,
R.sup.18 are the same as described before.
[0085] Step 1:
[0086] X.sup.1 is a leaving group such as halogen (Cl, Br, I), OTf,
OTs or OMs etc. Buchwald reaction is carried out in 1,4-dioxane or
N, N-dimethylacetamide. Base such as cesium carbonate or t-BuONa is
added. The catalyst used is tris(dibenzylideneacetone)dipalladium
or palladium acetate. The ligand used is
4,5-bisdiphenylphosphino-9,9-dimethyloxaxene or
2-(dicyclohexylphosphine)-3,6-dimethoxy-2',4',
6'-triisopropyl-1,1'-biphenyl. The reaction takes place at
110.about.150.degree. C. in an oil bath or under microwave
conditions to give compound (XIII). When X.sup.1 is Cl, the
substitution takes place in hydrochloric acid/1, 4-dioxane solution
in a sealed tube at 100.degree. C. to give Compound (XIII).
##STR00027##
[0087] The definitions of R.sup.2, R.sup.12, R.sup.16-R.sup.18 are
the same as described before.
[0088] Step 1:
[0089] X.sup.1 is a leaving group such as halogen (Cl, Br, I), OTf,
OTs, OMs etc. Buchwald reaction is carried out in 1,4-dioxane or N,
N-dimethylacetamide. At the same time, cesium carbonate or sodium
t-butoxide is added as a base, and the catalyst used is
tris(dibenzylideneacetone)dipalladium or palladium acetate. The
ligand used is 4,5-bisdiphenylphosphine-9,9-dimethyl xanthene or
2-(dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphen-
yl, etc. The reaction takes place at 80.about.110.degree. C. in an
oil bath or under microwave conditions to give compound (XIII).
##STR00028##
[0090] The definitions of R.sup.12, R.sup.16, R.sup.17, R.sup.20
are the same as described before.
[0091] Step 1:
[0092] X.sup.1 is a leaving group such as halogen (Cl, Br, I), OTf,
OTs, OMs etc. L is selected from O or NH. Buchwald reaction is
carried out in solvents such as 1,4-dioxane or N,
N-dimethylacetamide. At the same time, cesium carbonate or sodium
t-butoxide is added as a base, and the catalyst used is
tris(dibenzylideneacetone)dipalladium or palladium acetate. The
ligand used is 4,5-bisdiphenylphosphine-9,9-dimethyl xanthene or
2-(dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphen-
yl, etc. The reaction takes place at 110.about.150.degree. C. in an
oil bath or under microwave conditions to give Compound (XIV).
DETAILED DESCRIPTION OF THE INVENTION
Definition
[0093] Unless stated to the contrary, the following terms used in
the description and the claims have the following meanings.
[0094] The expression "Cx-Cy" as used herein represents the range
of the number of carbon atoms, where both x and y are integers. For
example, C3-C8 cyclyl represents a cyclyl group having 3 to 8
carbon atoms, and --C0-C2 alkyl represents an alkyl group having 0
to 2 carbon atoms, where --CO alkyl refers to a single chemical
bond.
[0095] The term "alkyl" refers to a saturated aliphatic hydrocarbon
group, including linear and branched groups having 1 to 20 carbon
atoms, for example, linear and branched groups having 1 to 18
carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6
carbon atoms or 1 to 4 carbon atoms. Non-limiting examples include
methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl,
s-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl,
2,2-dimethyl propyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl,
n-hexyl, 1-ethyl-2-methyl propyl, 1,1,2-trimethylpropyl,
1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethyl butyl,
1,3-dimethylbutyl, 2-ethylbutyl, and various branched isomers
thereof, etc. Alkyl may be substituted or unsubstituted.
[0096] The term "cyclyl" or "cyclic group" refers to saturated or
partially unsaturated monocyclic or polycyclic hydrocarbon groups,
comprising 3 to 12 cyclic carbon atoms, such as 3 to 12, 3 to 10, 3
to 8 or 3 to 6 cyclic carbon atoms, or 3, 4, 5, 6-membered rings.
Non-limiting examples of monocyclic cyclyl include cyclopropyl,
cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl,
cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl and the
like. Cyclyl may be substituted or unsubstituted.
[0097] The term "heterocyclyl" refers to a saturated or partially
unsaturated monocyclic or polycyclic hydrocarbon group, comprising
3 to 20 ring atoms, such as 3 to 16, 3 to 12, 3 to 10, 3 to 8 or 3
to 6 ring atoms, where one or more ring atoms are heteroatoms
selected from the group consisting of nitrogen, oxygen or S(O)m
(where m is an integer of 0 to 2), but excluding ring parts of
--O--O--, --O--S-- or --S--S--, and the remaining ring atoms are
carbon. Preferably 3 to 12 ring atoms, of which 1 to 4 are
heteroatoms, are comprised. More preferably the heterocyclyl ring
comprises 3 to 10 ring atoms, more preferably 3 to 8 ring atoms.
Most preferred are 5-membered rings or 6-membered rings, where 1 to
4 members are heteroatoms, more preferably 1 to 3 are heteroatoms,
and most preferably 1 to 2 are heteroatoms. Non-limiting examples
of monocyclic heterocyclyl include pyrrolidinyl, piperidyl,
piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl and the
like. Polycyclic heterocyclic groups include spirocyclic, fused and
bridged cyclic heterocyclyl groups.
[0098] The term "spiroheterocyclic group" refers to a 5 to 20
membered polycyclic heterocyclic group with one atom (referred to
as a spiro atom) shared between monocyclic rings, where one or more
of the ring atoms are heteroatoms selected from the group
consisting of nitrogen, oxygen or S(O)m (where m is an integer of 0
to 2), and the rest of the ring atoms are carbon. They may contain
one or more double bonds, but none of the rings has a completely
conjugated .pi. electron system. They are preferably 6 to 14
membered, and more preferably 7 to 10 membered. According to the
number of spiro atoms shared between rings, spirocyclyl groups are
divided into mono-spiroheterocyclyl, bi-spiroheterocyclyl or
poly-spiroheterocyclyl, preferably mono-spirocyclyl and
bi-spirocyclyl, and more preferably 4 membered/4 membered, 4
membered/5 membered, 4 membered/6 membered, 5 membered/5 membered,
or 5 membered/6 membered mono-spirocyclyl. Non-limiting examples of
spirocyclyl include
##STR00029##
[0099] The term "fused heterocyclyl" refers to a 5 to 20 membered
polycyclic heterocyclyl group where each ring in the system shares
a pair of adjacent atoms with other rings in the system, one or
more rings may contain one or more double bonds, but none of the
rings has a completely conjugated pi electron system, where one or
more ring atoms are heteroatoms selected from the group consisting
of nitrogen, oxygen or S(O)m (where m is an integer of 0 to 2), and
the remaining ring atoms are carbon. They are preferably 6 to 14
membered, and more preferably 7 to 10 membered. According to the
number of rings, they can be divided into bicyclic, tricyclic,
tetracyclic or polycyclic fused heterocyclyl, and the fused
heterocyclyl groups are preferably bicyclic or tricyclic, and more
preferably 5 membered/5 membered, or 5 membered/6 membered bicyclic
fused heterocyclyl. Non-limiting examples of fused heterocyclyl
include
##STR00030##
[0100] The heterocyclyl ring may be fused to an aryl, a heteroaryl
or a cyclyl ring, in which the ring connected with the parent
structure is a heterocyclyl group, and the non-limiting examples
include:
##STR00031##
and the like.
[0101] The heterocyclyl group may be substituted or
unsubstituted.
[0102] The term "aryl" refers to a 6 to 14 membered all-carbon
monocyclic or condensed polycyclic (i.e., rings sharing adjacent
pairs of carbon atoms) group, and a polycyclic (i.e., rings bearing
adjacent pairs of carbon atoms) group having a conjugated
pi-electron system, preferably 6 to 10 membered, for example,
phenyl and naphthyl, and most preferably phenyl. The aryl ring may
be fused to a heteroaryl, a heterocyclyl or a cyclyl ring, in which
the ring connected with the parent structure is an aryl ring, and
the non-limiting examples include:
##STR00032##
[0103] Aryl may be substituted or unsubstituted.
[0104] The term "heteroaryl" herein refers to a heteroaromatic
system comprising 1 to 4 heteroatoms and 5 to 14 ring atoms, where
the heteroatoms include oxygen, sulfur and nitrogen. Heteroaryl is
preferably 5 to 10 membered, and more preferably 5 membered or 6
membered, e.g., furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl,
pyrimidinyl, pyrazinyl, imidazolyl, tetrazyl, oxazolyl, and
isoxazolyl etc. The heteroaryl ring can be fused to an aryl, a
heterocyclyl or a cyclyl ring, where the ring connected with the
parent structure is a heteroaryl ring, and the non-limiting
examples include:
##STR00033##
[0105] Heteroaryl may be substituted or unsubstituted.
[0106] The term "halogen" herein refers to fluorine, chlorine,
bromine or iodine.
[0107] The term "cyano" herein refers to --CN.
[0108] The term "alkenyl" refers to a linear, branched hydrocarbon
group containing at least one carbon-carbon double bond, including
linear and branched groups having 2 to 20 carbon atoms, for
example, linear and branched groups having 2 to 18 carbon atoms, 2
to 12 carbon atoms, 2 to 8 carbon atoms, 2 to 6 carbon atoms or 2
to 4 carbon atoms. Where 1 to 3 carbon-carbon double bonds may be
present and preferably 1 carbon-carbon double bond may be present.
The term "C2-4 alkenyl" refers to alkenyl having 2 to 4 carbon
atoms, including vinyl, propenyl, butenyl, 2-methylbutenyl. The
alkenyl group may optionally be substituted.
[0109] The term "alkynyl" refers to a linear, or branched
hydrocarbon group containing at least one carbon-carbon triple
bond, including linear and branched groups having 2 to 20 carbon
atoms, for example, linear and branched groups having 2 to 18
carbon atoms, 2 to 12 carbon atoms, 2 to 8 carbon atoms, 2 to 6
carbon atoms or 2 to 4 carbon atoms. Among them, 1 to 3
carbon-carbon triple bonds may be present and preferably 1
carbon-carbon triple bond may be present. The term "C2-4 alkynyl"
refers to alkynyl having 2 to 4 carbon atoms, Non-limiting examples
including acetenyl, propynyl, butynyl and 3-methyl-1-butynyl.
[0110] The term "heteroalkyl" refers to a stable straight-chain or
branched-chain hydrocarbon group consisting of a specified number
of carbon atoms and at least one heteroatom selected from oxygen,
nitrogen and sulfur. Among them, nitrogen and sulfur atoms may be
oxidized optionally, nitrogen atoms may be quaternized optionally,
and hetero atoms such as oxygen, nitrogen and sulfur may be located
at any internal position of the heteroalkyl group, or at the
position where the alkyl group is connected with the rest of the
molecule. More than two heteroatoms may be independent or
continuous.
[0111] The term "alkyloxy" refers to the alkyl group connected by
an oxygen bridge, comprising alkyloxy group, cyclyloxy and
heterocyclyloxy group. Thus, the alkyl in the therm "alkoxy"
includes alkyl, heterocyclyl and cyclyl or cyclic group as defined
above.
[0112] The "optional" and "optionally" means that an event or
environment described subsequently may but does not necessarily
occur, including cases where the event or environment occurs or
does not occur. For example, "heterocyclyl optionally substituted
by alkyl" means that alkyl may but does not necessarily exist,
including cases where heterocyclyl is substituted by alkyl and not
substituted by alkyl.
[0113] The term "substituted" means that one or more hydrogen
atoms, preferably at most 5 and more preferably 1 to 3 hydrogen
atoms, in a group are substituted independently with corresponding
number of substituents. It goes without saying that, substituents
are only located in their possible chemical positions, and a person
skilled in the art can determine (experimentally or theoretically)
possible or impossible substitutions without a lot of efforts. For
example, amino or hydroxy groups having free hydrogen may be
unstable when combined with carbon atoms having unsaturated (e.g.
olefinic) bonds.
[0114] The substituent(s) include, but are not limited to, the
alkyl, alkenyl, alkynyl, alkoxy, halogen, hydroxyl, amino, cyano
and thiol groups.
[0115] The term "pharmaceutical composition" represents a mixture
of one or more of the compounds described herein or
physiologically/pharmaceutically acceptable salts or prodrugs with
other chemical components, as well as other components such as
physiologically/pharmaceutically acceptable carriers and
excipients. An object of the pharmaceutical compositions is to
promote the administration of drugs to organisms, facilitate the
absorption of active ingredients and thus exert biological
activity.
[0116] The term "room temperature" refers to 15 to 30.degree.
C.
[0117] The term "a stable isotopic derivative" includes:
derivatives substituted with isotopes, such as derivatives obtained
by substituting any hydrogen atom in Formula I with 1 to 5
deuterium atoms, derivatives substituted with isotopes obtained by
substituting any carbon atom in Formula I with 1 to 3 carbon-14
atoms, or derivatives substituted with isotopes obtained by
substituting any oxygen atom in Formula I with 1 to 3 .sup.18O
atoms.
[0118] The "pharmaceutically acceptable salts" as described in the
present invention are discussed in Berge, et al., "Pharmaceutically
acceptable salts," J. Pharm. Sci., 66, 1-19 (1977), and it is
obvious to pharmaceutical chemists that said salts are essentially
non-toxic and can provide desired pharmacokinetic properties,
palatability, absorption, distribution, metabolism or excretion,
and the like.
[0119] The pharmaceutically acceptable salts according to the
present invention can be synthesized through a general chemical
method.
[0120] In general, the preparation of the salts can be achieved by
reacting the compounds in the form of free alkalis or acids with
equivalent chemical equivalents or excess amounts of acids
(inorganic or organic acids) or alkalis in suitable solvents or
solvent compositions.
[0121] The "prodrug" as described in the present invention refers
to a compound that can be converted into an original active
compound after being metabolized in vivo. Representatively
speaking, prodrugs are inactive substances, or have activity lower
than the active parent compounds but can provide convenient
operation and dosage or improve metabolic characteristics.
[0122] The "isomer" of the present invention means that the
compound of Formula (I) according to the present invention may have
one or more asymmetric center and may be a racemate, a racemic
mixture and a single diastereoisomer. The isomers such as
stereoisomers and geometric isomers are all included in the present
invention. The geometric isomers include cis- and
trans-isomers.
[0123] The invention shall include any polymorph of the compound or
its salts, as well as any kind of hydrate or other solvate.
EXAMPLES
[0124] The present invention will be further illustrated by means
of examples below, but is not indented to limited to the scope of
the examples described. In the following examples, experimental
methods without specific conditions noted are selected according to
conventional methods and conditions or according to product
instructions.
[0125] The structures of all the compounds according to the present
invention can be identified by nuclear magnetic resonance (.sup.1H
NMR) and/or mass spectrometric detection (MS).
[0126] .sup.1H NMR chemical shift (5) is recorded in PPM (unit:
10.sup.-6 PPM). NMR is carried out by a Bruker AVANCE-400
spectrometer. Appropriate solvents include deuterated chloroform
(CDCl.sub.3), deuterated methanol (CD.sub.3OD) and deuterated
dimethylsulfoxide (DMSO-d.sup.6), with tetramethylsilane (TMS) as
an internal standard.
[0127] The low resolution mass spectrogram (MS) is determined by an
Agilent 1260HPLC/6120 mass spectrometer, using Agilent ZORBAX
XDB-C18, 4.6.times.50 mm, 3.5 .mu.m, at a gradient elution
condition I: 0:95% solvent A1 and 5% solvent B1, 1-2:5% solvent A1
and 95% solvent B1; 2.01-2.50: 95% solvent A1 and 5% solvent B1.
The percentage is the volume percentage of a certain solvent based
on the total solvent volume. Solvent A1: 0.01% formic acid aqueous
solution; solvent B1: 0.01% formic acid solution in acetonitrile;
and the percentage is the volume percentage of a solute based on
the solution.
[0128] The thin-layer silica gel plate is a Yantai Yellow Sea
HSGF254 or Qingdao GF254 silica gel plate. The Yantai Yellow Sea
100-200 or 200-300 mesh silica gel is generally used as the support
in the column chromatography.
[0129] The prep-HPLC uses the Waters SQD2 Ms guided HPLC,
XBridge-C18, 30.times.150 mm preparation column, 5 um. Method 1:
acetonitrile-water (0.2% formic acid), flow rate: 25 ml/min, Method
II: acetonitrile-water (0.8% ammonium bicarbonate), flow rate: 25
ml/min.
[0130] The known starting raw materials of the present invention
can be synthesized by or in accordance with methods known in the
art, or can be purchased from companies such as Acros Organics,
Aldrich Chemical Company, Accela ChemBio Inc., Shanghai Bide
Pharmatech, Shanghai Aladdin Chemistry, Shanghai Meryer Chemistry,
Accelerating Chemistry, Energy Chemistry etc.
[0131] In the examples, unless otherwise specified, the solvents
used in the reaction are all anhydrous solvents, where anhydrous
tetrahydrofuran is commercially available tetrahydrofuran, sodium
blocks are used as a dehydrant, benzophenone is used as an
indicator, the solution is refluxed under the protection of argon
until the it maintains a blue violet color, it is distilled and
collected, and stored at room temperature under the protection of
argon, and the other anhydrous solvents are purchased from Energy
Chemistry and Accelerating Chemistry, and the transfer and use of
all anhydrous solvents shall be carried out under the protection of
argon Unless otherwise specified.
[0132] In the examples, the reactions are all carried out under an
argon atmosphere or nitrogen atmosphere unless otherwise
specified.
[0133] The argon atmosphere or nitrogen atmosphere means that the
reaction flask is connected to an argon or nitrogen balloon with a
volume of about 1 L.
[0134] The hydrogen atmosphere means that the reaction flask is
connected to a hydrogen balloon with a volume of about 1 L.
[0135] In hydrogenation, the reaction is usually vacuumed and
filled with hydrogen gas, and this procedure is repeated for 3
times.
[0136] The reaction temperature is the room temperature, and the
temperature range is from 15.degree. C. to 30.degree. C., unless
otherwise specified.
[0137] The thin-layer chromatography method (TLC) is employed to
monitor the reaction process in the examples. The developer system
used in the reaction includes: A, a dichloromethane and methanol
system, and B: a petroleum ether and ethyl acetate system, and the
ratio by volume of the solvents is adjusted according to the
polarity of the compounds.
[0138] The eluent system for column chromatography and the
developer system for thin-layer chromatography employed in the
purification of compounds include: A, a dichloromethane and
methanol system, and B: a petroleum ether and ethyl acetate system,
and the ratio by volume of the solvents is adjusted according to
the polarity of the compounds, and a small amount of triethyl amine
and acid or alkaline reagents and the like can also be added for
the adjustment.
Example 1
1-tert-Butoxycarbonyl-5-(6-acetylaminopyrimidin-4-ylamino)-6-methoxyindazo-
le
##STR00034##
[0140] Step 1
1-tert-Butoxycarbonyl-5-bromo-6-methoxyindazole
[0141] 5-bromo-6-methoxy-1H-indazole 1a (4 g, 17.6 mmol),
triethylamine (5.3 g, 52.8 mmol), di-t-butyl dicarbonate (7.7 g,
35.2 mmol), 4-dimethylaminopyridine (7.7 g, 35.2 mmol) and
tetrahydrofuran (40 mL) were mixed and stirred at room temperature
for 1 hour under argon. The mixture was concentrated under reduced
pressure. Purification of the residue using column chromatography
(petroleum ether/ethyl acetate=4/1) gave
1-tert-butoxycarbonyl-5-bromo-6-methoxyindazole 1b (2.8 g, 8.59
mmol, yellow solid). Yield: 49%.
[0142] MS m/z (ESI): 327 & 329 [M+1],
[0143] Step 2
1-tert-Butoxycarbonyl-5-(6-acetamidopyrimidin-4-ylamino)-6-methoxyindazole
[0144] To the mixture of
1-tert-butoxycarbonyl-5-bromo-6-methoxyindazole 1b (24.0 mg, 0.08
mmol), N-(6-aminopyrimidin-4-yl)acetamide (12.0 mg, 0.08 mmol) and
1,4-dioxane (2.0 mL) were added
tris(dibenzylideneacetone)dipalladium (8.0 mg, 0.008 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (10.0 mg, 0.016
mmol) and cesium carbonate (52.0 mg, 0.16 mmol) at room temperature
under argon. The mixture was stirred at 110.degree. C. in microwave
oven for 1 hour under argon, diluted with dichloromethane (10 mL)
and filtered. The filtrate was concentrated under reduced pressure.
Purification of the residue using Prep TLC
(dichloromethane/methanol=20/1)
gavel-tert-butoxycarbonyl-5-(6-acetamidopyrimidin-4-ylamino)-6-methoxyind-
azole 1 (10 mg, 0.025 mmol, yellow solid). Yield: 31%.
[0145] MS m/z (ESI): 399 [M+1],
[0146] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.61 (s, 1H), 8.42
(s, 1H), 8.39 (s, 1H), 8.03 (s, 1H), 7.69 (s, 1H), 7.55 (s, 1H),
7.38 (s, 1H), 3.98 (s, 3H), 2.15 (s, 3H), 1.66 (s, 9H).
Example 2
5-(6-Acetylaminopyrimidin-4-ylamino)-6-methoxy-1H-indazole
##STR00035##
[0148] The mixture of
1-tert-butoxycarbonyl-5-(6-acetylaminopyrimidin-4-ylamino)-6-methoxyindaz-
ole 2a (10 mg, 0.025 mmol) in dichloromethane (2.0 mL) and
trifluoroacetic acid (1.0 mL) was stirred at room temperature for 3
hours. The mixture was quenched with saturated sodium bicarbonate
aqueous solution (5 mL) and diluted with dichloromethane (5 mL).
The organic layer was separated and the aqueous layer was extracted
with dichloromethane (10 mL.times.2). The combined organic layer
was washed with brine (20 mL.times.2), dried over anhydrous sodium
sulfate and then filtered. The filtrate was concentrated under
reduced pressure. Purification of the residue by Prep-TLC
(dichloromethane/methanol=10/1) gave
5-(6-acetylaminopyrimidin-4-ylamino)-6-methoxy-1H-indazole 2 (5 mg,
0.017 mmol, yellow solid). Yield: 68%.
[0149] MS m/z (ESI): 299 [M+1],
[0150] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.83 (s, 1H), 10.34
(s, 1H), 8.73 (s, 1H), 8.26 (s, 1H), 7.94 (s, 1H), 7.93 (s, 1H),
7.35 (s, 1H), 7.03 (s, 1H), 3.85 (s, 3H), 3.17 (s, 3H).
Example 3
1-tert-Butoxycarbonyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxy-
indazole
##STR00036##
[0152] See example 1 for the synthetic procedures.
N-(6-aminopyrimidin-4-yl)cyclopropanecarboxamide is used instead of
N-(6-aminopyrimidine-4-yl)ethanamide to give
1-tert-butoxycarbonyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methox-
yindazole 3 (1.0 g, 2.4 mmol, yellow solid). Yield: 80%.
[0153] MS m/z (ESI): 425 [M+1],
[0154] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.78 (s, 1H), 8.70
(s, 1H), 8.51 (s, 1H), 8.11 (s, 1H), 7.75 (s, 1H), 7.62 (s, 1H),
7.46 (s, 1H), 4.03 (s, 3H) 1.73 (s, 9H), 1.60-1.52 (m, 1H),
0.95-0.93 (m, 4H).
Example 4
5-(6-Cyclopropionamidopyrimidin-4-ylamino)-6-methoxy-1H-indazole
##STR00037##
[0156] See example 2 for the synthetic procedures.
1-tert-butoxycarbonyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methox-
yindazole is used instead of
1-tert-butoxycarbonyl-5-(6-acylamidopyrimidin-4-ylamino)-6-methoxyindazol-
e to give
5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxy-1H-indazole 4
(5.0 mg, 0.017 mmol, yellow solid). Yield: 68%.
[0157] MS m/z (ESI): 325 [M+1],
[0158] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.83 (s, 1H), 10.34
(s, 1H), 8.73 (s, 1H), 8.26 (s, 1H), 7.94 (s, 1H), 7.93 (s, 1H),
7.35 (s, 1H), 7.03 (s, 1H), 3.85 (s, 3H), 1.98-1.96 (m, 1H),
0.78-0.76 (m, 4H).
Example 5
5-(6-Cyclopropionamidopyrimidin-4-ylamino)-6-methoxyl-methylindazole
formate
##STR00038##
[0160] Step 1
5-Bromo-6-methoxy-1-methylindazole
[0161] To the mixture of 5-bromo-6-methoxy-1H-indazole 5a (113.0
mg, 0.5 mmol), cesium carbonate (326.0 mg, 1.0 mmol) and
N,N-dimethylacetamide (10 mL) was added iodomethane (84.0 mg, 0.6
mmol) at room temperature. The reaction mixture was stirred at room
temperature for 15 hours. It was quenched with saturated aqueous
sodium bicarbonate (10 mL) and extracted with dichloromethane (30
mL.times.3). The combined organic layer was dried over anhydrous
sodium sulfate and filtered. The filtrate was concentrated.
Purification of the residue using Prep-TLC (petroleum ether/ethyl
acetate=2/1) gave 5-bromo-6-methoxy-1-methylindazole 5b (15 mg,
0.062 mmol, yellow solid). Yield: 12%. MS m/z (ESI): 241 & 243
[M+1],
[0162] Step 2
[0163]
5-(6-Cyclopropionamidopyrimidin-4-ylamino)-6-methoxyl-methylindazol-
e To the mixture of 5-bromo-6-methoxy-1-methylindazole 5b (15.0 mg,
0.062 mmol), N-(6-aminopyrimidin-4-yl)cyclopropanecarboxamide (12.0
mg, 0.08 mmol) and dioxane (2.0 mL) were added
tris(dibenzylideneacetone)dipalladium (5.0 mg, 0.005 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (6.0 mg, 0.01 mmol)
and cesium carbonate (98.0 mg, 0.3 mmol) under argon. The mixture
was reacted at 110.degree. C. in microwave oven for 1 hour under
argon. It was cooled to room temperature, diluted with
dichloromethane (10 mL) and filtered. The filtrate was concentrated
under reduced pressure. Purification of the residue using Prep-HPLC
(water (0.2% formic acid), 10%.about.40% acetonitrile, 15 minutes)
gave
5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxyl-methylindazole
formate 5 (6.0 mg, 0.018 mmol, yellow solid). Yield: 29%.
[0164] MS m/z (ESI): 339 [M+1],
[0165] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.66 (s, 1H), 8.71
(s, 1H), 8.29 (s, 1H), 8.27 (s, 1H), 7.91 (s, 1H), 7.90 (s, 1H),
7.30 (s, 1H), 7.21 (s, 1H), 4.02 (s, 3H), 3.88 (s, 3H), 2.01-1.92
(m, 1H), 0.80-0.77 (m, 4H).
Example 6
5-(6-Cyclopropionamidopyrimidin-4-ylamino)-6-ethoxy-1H-indazole
##STR00039##
[0167] Step 1
5-Bromo-1H-indazol-6-ol
[0168] 5-Bromo-6-methoxy-1H-indazole 6a (450.0 mg, 2.0 mmol) was
mixed with aqueous solution of hydrobromic acid (10 mL) and the
mixture was stirred at 100.degree. C. for 15 hours. It was cooled
to room temperature, quenched with saturated aqueous sodium
bicarbonate (20 mL) and extracted with dichloromethane (30
mL.times.3). The combined organic layer was dried over anhydrous
sodium sulfate and then filtered. The filtrate was concentrated
under reduced pressure. Purification of the residue using Prep-TLC
(petroleum ether/ethyl acetate=1/1) gave 5-bromo-1H-indazol-6-ol 6b
(200.0 mg, 1.2 mmol, yellow solid). Yield: 61%.
[0169] MS m/z (ESI): 213 & 215 [M+1],
[0170] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.70 (s, 1H), 10.35
(s, 1H), 7.91 (s, 1H), 7.87 (s, 1H), 6.99 (s, 1H).
[0171] Step 2
5-Bromo-6-ethoxy-1H-indazole
[0172] To the mixture of 5-bromo-1H-indazol-6-ol 6b (42.0 mg, 0.2
mmol), potassium carbonate (138.0 mg, 1.0 mmol) and acetone (5 mL)
was added iodoethane (50.0 mg, 0.3 mmol) at room temperature. The
mixture was stirred at room temperature for 3 hours. It was
quenched with saturated aqueous sodium bicarbonate (10 mL) and
extracted with ethyl acetate (10 mL.times.3). The combined organic
layer was dried over anhydrous sodium sulfate and then filtered.
The filtrate was concentrated under reduced pressure. Purification
of the residue using Prep-TLC (petroleum ether/ethyl acetate=1/1)
gave 5-bromo-6-ethoxy-1H-indazole 6c (10.0 mg, 0.041 mmol, white
solid). Yield: 21%.
[0173] MS m/z (ESI): 241 & 243 [M+1],
[0174] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.14 (s, 1H),
7.86 (s, 1H), 7.85 (s, 1H), 6.81 (s, 1H), 4.08-4.02 (m, 2H), 1.46
(t, J=7.6 Hz, 3H).
[0175] Step 3
1-t-Butoxycarbonyl-5-bromo-6-ethoxyindazole
[0176] To the mixture of 5-bromo-6-ethoxy-1H-indazole 6c (10.0 mg,
0.04 mmol), triethylamine (10.0 mg, 0.1 mmol) and tetrahydrofuran
(2 mL) was added di-t-butyldicarbonate (8.0 mg, 0.04 mmol) at room
temperature. The reaction mixture was stirred at room temperature
for 3 hours. It was quenched with saturated aqueous sodium
bicarbonate (10 mL) and extracted with ethyl acetate (10
mL.times.3). The combined organic layer was dried over anhydrous
sodium sulfate and then filtered. The filtrate was concentrated
under reduced pressure. Purification of the residue using Prep-TLC
(petroleum ether/ethyl acetate=5/1) gave
1-t-Butoxycarbonyl-5-bromo-6-ethoxyindazole 6d (10.0 mg, 0.03 mmol,
white solid).
[0177] Yield: 75%. MS m/z (ESI): 341 & 343 [M+1],
[0178] Step 4
1-t-Butoxycarbonyl-5-(6-cyclopropanecarboxamido)pyrimidin-4-ylamino)-6-eth-
oxyindazole
[0179] To the mixture of
1-t-Butoxycarbonyl-5-bromo-6-ethoxyindazole 6d (10.0 mg, 0.03
mmol), N-(6-aminopyrimidin-4-yl)cyclopropanecarboxamide (5.0 mg,
0.03 mmol) and 1,4-dioxane (1.0 mL) were added
tris(dibenzylideneacetone)dipalladium(0) (3.0 mg, 0.003 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (6.0 mg, 0.01 mmol)
and cesium carbonate (32.0 mg, 0.1 mmol) under argon. The mixture
was stirred at 110.degree. C. in microwave oven for 1 hour under
argon and cooled to room temperature. The mixture was diluted with
dichloromethane (10 mL) and filtered. The filtrate was concentrated
under reduced pressure. Purification of the residue using Prep-TLC
(dichloromethane/methanol=20/1) gave
1-t-Butoxycarbonyl-5-(6-cyclopropanecarboxamidopyrimidin-4-ylamino)-6-eth-
oxyindazole 6e (4.5 mg, 0.01 mmol, white solid). Yield: 33%.
[0180] MS m/z (ESI): 439 [M+1].
[0181] Step 5
5-(6-Cyclopropionamidopyrimidin-4-ylamino)-6-ethoxy-1H-indazole
[0182]
1-t-Butoxycarbonyl-5-(6-cyclopropanecarboxamidopyrimidin-4-ylamino)-
-6-ethoxyindazole 6e (4.5 mg, 0.01 mmol), dichloromethane (1.0 mL)
and trifluoroacetic acid (1.0 mL) were mixed and stirred at room
temperature for 3 hours. The mixture was quenched with saturated
aqueous sodium bicarbonate (5 mL) and diluted with dichloromethane
(5 ml). The organic layer is separated, and the aqueous layer is
extracted with dichloromethane (5 mL). The combined organic layer
was washed with brine (20 mL.times.2), dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated under reduced
pressure. Purification of the residue using Prep-TLC
(dichloromethane/methanol=10:1) gave
5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-ethoxy-1H-indazole 6
(2.0 mg, 0.06 mmol, white solid). Yield: 60%.
[0183] MS m/z (ESI): 339 [M+1],
[0184] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.66 (s, 1H), 8.62
(s, 1H), 8.27 (s, 1H), 8.23 (s, 1H), 7.93 (s, 1H), 7.88 (s, 1H),
7.30 (s, 1H), 7.01 (s, 1H), 4.12-4.07 (m, 2H), 2.01-1.92 (m, 1H),
1.32-1.28 (d, J=4.8 Hz, 3H), 0.80-0.78 (m, 4H).
Example 7
5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-isopropoxy-1H-indazole
formate
##STR00040##
[0186] With similar procedures to those in Example 6,
2-bromopropane is used instead of iodoethane, the desired compound
7:
5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-isopropoxy-1H-indazole
formate (1.3 mg, 0.004 mmol, white solid) was obtained. Yield:
40%.
[0187] Prep HPLC condition: (water (0.2% formic acid),
15%.about.35% acetonitrile, 15 minutes)
[0188] MS m/z (ESI): 352 [M+1],
[0189] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.77 (s, 1H), 10.65
(s, 1H), 8.54 (s, 1H), 8.28 (s, 1H), 8.24 (s, 1H), 7.93 (s, 1H),
7.89 (s, 1H), 7.30 (s, 1H), 7.02 (s, 1H), 4.68-4.62 (m, 1H),
1.99-1.96 (m, 1H), 1.25 (d, J=5.6 Hz, 6H), 0.77-0.75 (m, 4H).
Example 8
1-Cyclohexanoyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxyindazo-
le
##STR00041##
[0191] To the mixture of
5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxy-1H-indazole 8a
(10.0 mg, 0.03 mmol), sodium hydride (60% dispersion in mineral
oil, 3.0 mg, 0.1 mmol) and tetrahydrofuran (2.0 mL) was added
cyclohexanecarbonyl chloride (6.0 mg, 0.03 mmol). The mixture was
reacted at room temperature for 10 minutes. The mixture was
quenched with saturated aqueous sodium bicarbonate (10 mL) and
extracted with dichloromethane (10 mL.times.3). The combined
organic layer was dried over anhydrous sodium sulfate and filtered.
The filtrate was concentrated under reduced pressure. Purification
of the residue using Prep-TLC (dichloromethane/methanol=10/1) gave
1-cyclohexanoyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxyindaz-
ole 8 (2.0 mg, 0.005 mmol, yellow solid). Yield: 15%.
[0192] MS m/z (ESI): 435 [M+1],
[0193] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.88 (s, 1H), 9.16
(s, 1H), 8.54 (s, 1H), 8.12 (s, 1H), 7.97 (s, 1H), 7.06 (s, 1H),
6.65 (s, 1H), 3.88 (s, 3H), 2.84-2.81 (m, 1H), 2.03-2.00 (m, 2H),
1.89-1.86 (m, 2H), 1.80-1.77 (m, 3H), 1.73-1.71 (m, 1H), 1.46-1.42
(m, 2H), 0.97-0.94 (m, 4H), 0.88-0.84 (m, 1H).
Example 9
5-(6-aminopyrimidin-4-ylamino)-6-methoxy-1H-indazole
##STR00042##
[0195] To the mixture of
1-tert-butoxycarbonyl-5-bromo-6-methoxyindazole 9a (50.0 mg, 0.15
mmo), 4,6-diaminopyrimidine (22.0 mg, 0.2 mmol) and 1,4-dioxane
(1.0 mL) was added tris(dibenzylideneacetone)dipalladium(0) (14.0
mg, 0.015 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene
(17.0 mg, 0.03 mmol) and cesium carbonate (98.0 mg, 0.3 mmol) under
argon. The mixture was reacted at 125.degree. C. in microwave oven
for 1 hour and cooled to room temperature. The mixture was diluted
with dichloromethane (10 mL) and filtered, the filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep-TLC (dichloromethane/methanol=10/1) gave
5-(6-aminopyrimidin-4-ylamino)-6-methoxy-1H-indazole 9 (2.0 mg,
0.01 mmol, white solid). Yield: 7%.
[0196] MS m/z (ESI): 257 [M+1],
[0197] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.28 (s, 1H), 7.96
(s, 1H), 7.92 (s, 1H), 7.87 (s, 1H), 7.84 (s, 1H), 7.02 (s, 1H),
6.17 (s, 2H), 5.58 (s, 1H), 3.86 (s, 3H).
Example 10
5-(6-benzoylaminopyrimidin-4-ylamino)-6-methoxy-1H-indazole
##STR00043##
[0199] The synthetic procedure is similar to that in Example 1.
using N-(6-aminopyrimidine-4-yl)benzamide instead of
N-(6-aminopyrimidine-4-yl)ethanamide, The desired compound 10
5-(6-benzoylaminopyrimidin-4-ylamino)-6-methoxy-1H-indazole was
obtained. Yield: 42%.
[0200] MS m/z (ESI): 361 [M+1],
[0201] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.88 (s, 1H), 10.69
(s, 1H), 8.86 (s, 1H), 8.35 (s, 1H), 8.11-7.89 (m, 4H), 7.70-7.35
(m, 4H), 7.05 (s, 1H), 3.87 (s, 3H).
Example 11
5-(6-(2-Pyridine)
formylaminopyrimidin-4-ylamino)-6-methoxy-1H-indazole
##STR00044##
[0203] Step 1
5-Bromo-6-methoxy-1-((2-(trimethylsilyl) ethoxy)
methyl)-1H-indazole
[0204] 5-Bromo-6-methoxy-1H-indazole 11a (320 mg, 1.4 mmol),
N,N-diisopropylethylamine (903 mg, 7 mmol), 2-(trimethylsilyl)
ethoxymethyl chloride (500 mg, 3.0 mmol) and dichloromethane (10
mL) were mixed and reacted at room temperature for 3 hours under
argon. The mixture was concentrated under reduced pressure.
Purification of the residue using column chromatography (petroleum
ether/ethyl acetate=4/1) gave
5-bromo-6-methoxy-1-((2-(trimethylsilyl) ethoxy)
methyl)-1H-indazole 11 b (320 mg, 0.9 mmol, yellow solid).
[0205] Yield: 64%.
[0206] MS m/z (ESI): 357 & 359 [M+1],
[0207] Step 2
1-((2-(Trimethylsilyl) ethoxy) methyl)-5-(6-(2-pyridine)
formylamino) pyrimidin-4-ylamino)-6-methoxyindazole
[0208] 5-Bromo-6-methoxy-1-((2-(trimethylsilyl) ethoxy)
methyl)-1H-indazole 11b (18.0 mg, 0.05 mmol),
2-(7-azabenzotriazol-1-yl)-N,N,N',N'tetramethyluronium
hexafluorophosphate (38.0 mg, 0.10 mmol), N,N-diisopropylethylamine
(13.0 mg, 0.10 mmol) were dissolved into tetrahydrofuran (1 mL) and
stirred at room temperature for 15 minutes, then picolinic acid
(12.0 mg, 0.10 mmol) was added. The reaction mixture was stirred at
room temperature for 12 hours. The mixture was concentrated under
reduced pressure. Purification of the residue using Prep-HPLC
(water (0.2% formic acid), 30%-70% acetonitrile, 15 minutes)
gavel-((2-(trimethylsilyl) ethoxy) methyl)-5-(6-(2-pyridine)
formylamino) pyrimidin-4-ylamino)-6-methoxyindazole 11c (6.0 mg,
0.012 mmol, white solid). Yield: 24%.
[0209] MS m/z (ESI): 492 [M+1],
[0210] Step 3
5-(6-(2-pyridine)
formylaminopyrimidin-4-ylamino)-6-methoxy-1H-indazole
[0211] 1-((2-(trimethylsilyl) ethoxy) methyl)-5-(6-(2-pyridine)
formylamino) pyrimidin-4-ylamino)-6-methoxyindazole 11c (6.0 mg,
0.012 mmol) was dissolved in dichloromethane (1.0 mL) and
trifluoroacetic acid (1.0 mL) was added, the mixture was stirred at
room temperature for 1.5 hours. The mixture was concentrated under
reduced pressure. Purification of the residue using Prep-HPLC
(water (0.2% formic acid), 30%.about.70% acetonitrile, 15 minutes)
gave 5-(6-(2-pyridine)
formylaminopyrimidin-4-ylamino)-6-methoxy-1H-indazole 11 (3.0 mg,
0.008 mmol, white solid). Yield: 67%.
[0212] MS m/z (ESI): 362 [M+1],
[0213] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.93 (brs, 1H),
10.19 (s, 1H), 9.03 (s, 1H), 8.76 (s, 1H), 8.35 (s, 1H), 8.31 (s,
1H), 8.18-8.09 (m, 2H), 7.97 (s, 1H), 7.76-7.72 (m, 1H), 7.52 (s,
1H), 7.07 (s, 1H), 3.87 (s, 3H).
Example 12
5-(6-Pivaloylaminopyrimidin-4-ylamino)-6-methoxy-1H-indazole
##STR00045##
[0215] Step 1
1-((2-(Trimethylsilyl) ethoxy) methyl)-5-(6-pivaloylamino)
pyrimidin-4-ylamino)-6-methoxyindazole
[0216] 1-((2-(Trimethylsilyl) ethoxy)
methyl)-5-(6-aminopyrimidin-4-ylamino)-6-methoxyindazole 12a (100.0
mg, 0.26 mmol) and triethylamine (55.0 mg, 0.52 mmol) are dissolved
in anhydrous dichloromethane (4.0 mL), pivaloyl chloride (50.0 mg,
0.39 mmol) was added. The mixture was stirred at room temperature
for 30 minutes. It was quenched with saturated aqueous sodium
bicarbonate (0.5 mL). The mixture was concentrated under reduced
pressure. Purification of the residue using Prep-TLC
(dichloromethane/methanol=20:1) gavel-((2-(trimethylsilyl) ethoxy)
methyl)-5-(6-pivaloylamino) pyrimidin-4-ylamino)-6-methoxyindazole
12b (100.0 mg, 0.21 mmol, yellow solid). Yield: 81%.
[0217] MS m/z (ESI): 471 [M+1],
[0218] Step 2
5-(6-Pivaloylaminopyrimidin-4-ylamino)-6-methoxy-1H-indazole
[0219] 1-((2-(Trimethylsilyl) ethoxy) methyl)-5-(6-pivaloylamino)
pyrimidin-4-ylamino)-6-methoxyindazole 12b (50.0 mg, 0.10 mmol) was
dissolved in dichloromethane (1.0 mL), and trifluoroacetic acid
(1.0 mL) was added and stirred at room temperature for 30 minutes.
The mixture was concentrated under reduced pressure. Purification
of the residue using Prep-HPLC (water (0.2% formic acid),
30%.about.70% acetonitrile, 15 minutes) gave
5-(6-pivaloylaminopyrimidin-4-ylamino)-6-methoxy-1H-indazole 12
(20.0 mg, 0.06 mmol, white solid). Yield: 60%.
[0220] MS m/z (ESI): 341 [M+1],
[0221] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.52-8.43 (m, 3H),
8.21 (brs, 1H), 8.01 (s, 1H), 7.68 (s, 1H), 7.44 (s, 1H), 6.92 (s,
1H), 3.95 (s, 3H), 1.29 (s, 9H).
Example 13
1-Isobutyryl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxyindazole
##STR00046##
[0223] Step 1
5-Bromo-6-methoxy-1-isobutyrylindazole
[0224] To the mixture of 5-bromo-6-methoxy-1H-indazole 13a (113.0
mg, 0.5 mmol), triethylamine (101.0 mg, 1.0 mmol) and
dichlormethane (10 mL) was added isobutyryl chloride (84.0 mg, 0.6
mmol) at room temperature, and was reacted at room temperature for
1 hour. The mixture was quenched with saturated aqueous sodium
bicarbonate (10 mL) and extracted with dichlormethane (30
mL.times.3). The combined organic layer was washed with brinebrine
(30 mL), dried over anhydrous sodium sulfate and then filtered. The
mixture was concentrated under reduced pressure. Purification of
the residue using Prep-TLC (petroleum ether/ethyl acetate=5/1) gave
5-bromo-6-methoxy-1-isobutyrylindazole 13b (41 mg, 0.14 mmol,
yellow solid). Yield: 28%.
[0225] MS m/z (ESI): 297 & 299 [M+1],
[0226] Step 2
1-Isobutyryl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxyindazole
[0227] To the mixture of 5-bromo-6-methoxy-1-isobutyrylindazole 13b
(15.0 mg, 0.050 mmol),
N-(6-aminopyrimidin-4-yl)cyclopropanecarboxamide (11.0 mg, 0.060
mmol) and 1,4-dioxane (2 mL) were added
tris(dibenzylideneacetone)-dipalladium(0) (5.0 mg, 0.005 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (6.0 mg, 0.010
mmol) and cesium carbonate (49.0 mg, 0.150 mmol) under argon. The
mixture was stirred at 110.degree. C. in microwave oven for 1 hour
under argon and cooled to room temperature. the mixture was diluted
with dichloromethane (10 mL) and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep-TLC (dichlormethane/methanol=20/1) gave
1-isobutyryl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxyindazole
13 (7.0 mg, 0.018 mmol, yellow solid). Yield: 36%.
[0228] MS m/z (ESI): 395 [M+1],
[0229] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.87 (s, 1H), 8.69
(s, 1H), 8.50 (s, 1H), 8.08 (s, 1H), 8.01 (s, 1H), 7.63 (s, 1H),
7.46 (s, 1H), 4.01 (s, 3H), 4.01-3.94 (m, 1H), 2.06-1.98 (m, 1H),
1.36 (d, J=7.2 Hz, 6H), 1.16-1.08 (m, 2H), 0.99-0.89 (m, 2H).
Example 14
1-Cyclopropanoyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxyindaz-
ole
##STR00047##
[0231] The synthetic procedure is similar to that in Example 13.
using cyclopropionyl chloride instead of isobutyryl chloride, the
desired compound
14:1-cyclopropanoyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-
-methoxyindazole (3.0 mg, 0.008 mmol) was obtained. Yield: 27%.
[0232] MS m/z (ESI): 393 [M+1],
[0233] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.74 (s, 1H), 8.51
(s, 1H), 8.17 (s, 1H), 8.13 (s, 1H), 7.97 (s, 1H), 7.61 (s, 1H),
7.43 (s, 1H), 4.01 (s, 3H), 1.33-1.30 (m, 2H), 1.16-1.10 (m, 4H),
0.97-0.85 (m, 4H).
Example 15
2-Cyclopropanoyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxyindaz-
ole
##STR00048##
[0235] To the mixture of
5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxy-1H-indazole
15a (50.0 mg, 0.15 mmol), N,N-diisopropylethylamine (60.0 mg, 0.45
mmol) and N,N-dimethylformamide (4 mL) was added
cyclopropanecarbonyl chloride (48.0 mg, 0.45 mmol). The mixture was
stirred at room temperature for 12 hours and was quenched with
saturated aqueous sodium bicarbonate (10 mL), then extracted with
dichloromethane (10 mL.times.3). The combined organic layer was
dried over anhydrous sodium sulfate and then filtered. The filtrate
was concentrated under reduced pressure. Purification of the
residue using Prep-HPLC (water (0.2% formic acid), 30%.about.70%
acetonitrile, 15 minutes) gave
2-cyclopropanoyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxyinda-
zole 15 (15 mg, 0.04 mmol, white solid). Yield: 25%.
[0236] MS m/z (ESI): 393 [M+1],
[0237] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.82 (s, 1H),
8.90 (s, 1H), 8.80 (s, 1H), 8.43 (s, 1H), 8.33 (s, 1H), 7.70 (s,
1H), 7.06 (s, 1H), 3.93 (s, 3H), 3.30-3.28 (m, 1H), 2.03-2.02 (m,
1H), 1.24-1.21 (m, 4H), 0.86-0.84 (m, 4H). .sup.13C NMR (400 MHz,
CDCl.sub.3) .delta. 173.89, 162.40, 157.91, 157.08, 154.83, 149.33,
129.57, 121.81, 118.45, 110.82, 95.20, 94.32, 56.45, 14.76, 12.73,
11.94, 8.53.
Example 16
1-Acetyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxyindazole
##STR00049##
[0239] The synthetic procedure is similar to that in Example 13.
acetic anhydride instead of isobutyryl chloride, the desired
compound 16:
1-acetyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxyindazole
(6.0 mg, 0.016 mmol, yellow solid) was obtained. Yield: 32%.
[0240] MS m/z (ESI): 367 [M+1],
[0241] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.83 (s, 1H), 8.50
(s, 1H), 8.17 (s, 1H), 8.08 (s, 1H), 7.96 (s, 1H), 7.61 (s, 1H),
7.42 (s, 1H), 4.03 (s, 3H), 2.79 (s, 3H), 2.06-1.97 (m, 1H),
1.15-1.09 (m, 2H), 0.99-0.92 (m, 2H).
Example 17
1-Methoxyacyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxyindazole
##STR00050##
[0243] The synthetic procedure is similar to that in Example 13.
using methyl chloroformate instead of isobutyryl chloride acetic
anhydride, The desired
compound17:1-methoxyacyl-5-(6-cyclopropionamidopyrimidin-4-ylamin-
o)-6-methoxyindazole (2.0 mg, 0.005 mmol, yellow solid) was
obtained. Yield: 10%.
[0244] MS m/z (ESI): 383 [M+1],
[0245] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.74 (s, 1H), 8.50
(s, 1H), 8.14 (s, 1H), 8.12 (s, 1H), 7.77 (s, 1H), 7.61 (s, 1H),
7.40 (s, 1H), 4.13 (s, 3H), 4.03 (s, 3H), 2.06-1.99 (m, 1H),
1.16-1.08 (m, 2H), 0.99-0.91 (m, 2H).
Example 18
1-Ethoxyacyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxyindazole
##STR00051##
[0247] The synthetic procedure is similar to that in Example 13.
using ethyl chloroformate instead of isobutyryl chloride, the
desired compound 18:
1-ethoxyacyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxyinda-
zole 18 (5.0 mg, 0.013 mmol, yellow solid) was obtained. Yield:
26%.
[0248] MS m/z (ESI): 397 [M+1],
[0249] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.74 (s, 1H), 8.50
(s, 1H), 8.16 (s, 1H), 8.13 (s, 1H), 7.78 (s, 1H), 7.61 (s, 1H),
7.40 (s, 1H), 4.83 (q, J=7.2 HZ, 2H), 4.03 (s, 3H), 2.06-1.99 (m,
1H), 1.54 (t, J=7.2 HZ, 3H), 1.16-1.08 (m, 2H), 0.99-0.91 (m,
2H).
Example 19
N-(6-((6-Methoxy-1H-indazol-5-yl)amino)pyrimidin-4-yl)isobutyramide
##STR00052##
[0251] Step 1
N-(6-Aminopyrimidin-4-yl)isobutyramide
[0252] To the solution of 4,6-diamino pyrimidine 19a (550 mg, 5.0
mmol) in tetrahydrofuran (5 mL) were added isobutyryl chloride
(1.07 g, 10.0 mmol) and potassium carbonate (2.07 g, 15.0 mmol) at
room temperature. The reaction mixture was kept at room temperature
overnight and concentrated under reduced pressure. Purification of
the residue using flash column chromatography
(dichloromethane/methanol=1:0.about.10:1) gave
N-(6-Aminopyrimidin-4-yl)isobutyramide 19b (80.6 mg, 0.4 mmol,
white solid). Yield: 9%.
[0253] MS m/z (ESI): 181 [M+1],
[0254] Step 2
N-(6-((6-Methoxy-1H-indazol-5-yl)amino)pyrimidin-4-yl)isobutyramide
[0255] To the mixture of N-(6-aminopyrimidin-4-yl)isobutyramide 19b
(3.6 mg, 0.02 mmol), 5-bromo-6-methoxy-1H-indazole (6 mg, 0.02
mmol), cesium carbonate (20 mg, 0.06 mmol) and 1,4-dioxane (1 mL)
were added tris(dibenzylideneacetone)dipalladium(0) (2.2 mg, 0.002
mmol) and
2-(dicyclohexylphosphine)-3,6-dimethoxy-2'-4'-6'-tri-isopropyl-1,1'-biphe-
nyl(2.2 mg, 0.004 mmol) under argon. The mixture was stirred at
110.degree. C. in oil bath for 1 hour under argon. Then it was
cooled to room temperature, diluted with methanol (5 mL) and
filtered. The filtrate was concentrated under reduced pressure.
Purification of the residue using Prep-HPLC gave
N-(6-((6-methoxy-1H-indazol-5-yl)amino)pyrimidin-4-yl)isobutyramide
19 (2.0 mg, 0.006 mmol, white solid). Yield: 30%.
[0256] MS m/z (ESI): 327 [M+1],
[0257] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.87 (s, 1H), 10.32
(s, 1H), 8.73 (s, 1H), 8.26 (s, 1H), 7.95-7.91 (m, 2H), 7.34 (s,
1H), 7.03 (s, 1H), 3.83 (s, 3H), 2.05-1.90 (m, 1H), 1.03 (d, J=6.7
Hz, 6H).
Example 20
5-(6-(Pyridin-2-ylamino) pyrimidin-4-ylamino)-6-methoxy-1H-indazole
formate
##STR00053##
[0259] Step 1
1-((2-(Trimethylsilyl) ethoxy)
methyl)-5-(6-aminopyrimidin-4-ylamino)-6-methoxyindazole
[0260] To a mixture of 4,6-diaminopyrimidine 20a (11.0 mg, 0.1
mmol),
5-bromo-6-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole
(36.0 mg, 0.05 mmol) and 1,4-dioxane (2.0 mL) were added
tris(dibenzylideneacetone)dipalladium(0) (9.0 mg, 0.01 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (12.0 mg, 0.02
mmol) and cesium carbonate (65.0 mg, 0.2 mmol) at room temperature
under agon. The mixture was stirred at 120.degree. C. for 16 hours.
It was cooled to room temperature and quenched with water (10 mL).
The organic layer was separated and the aqueous phase was extracted
with dichloromethane (15 mL.times.3). The combined organic layer
was washed with brine (30 mL), dried over anhydrous sodium sulfate
and then filtered, the filtrate was concentrated under reduced
pressure. Purification of the residue using Prep TLC
(dichloromethane/methanol=20/1) gave 1-((2-(trimethylsilyl) ethoxy)
methyl)-5-(6-aminopyrimidin-4-ylamino)-6-methoxyindazole 20b (22.0
mg, 0.057 mmol, yellow solid). Yield: 57%.
[0261] MS m/z (ESI): 387 [M+1],
[0262] Step 2
1-((2-(Trimethylsilyl) ethoxy) methyl)-5-(6-(pyridin-2-ylamino)
pyrimidin-4-ylamino)-6-methoxyindazole
[0263] To the mixture of 1-((2-(trimethylsilyl) ethoxy)
methyl)-5-(6-aminopyrimidin-4-ylamino)-6-methoxyindazole 20b (11.0
mg, 0.03 mmol), 2-bromopyridine (5.0 mg, 0.05 mmol) and 1,4-dioxane
(1.0 mL) were added tris(dibenzylideneacetone)dipalladium(0) (9.0
mg, 0.01 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene
(12.0 mg, 0.02 mmol) and cesium carbonate (65.0 mg, 0.2 mmol) at
room temperature under argon. The mixture was stirred at
120.degree. C. in microwave oven under argon for 1 hour, and then
was cooled to room temperature, quenched with water (10 mL). The
organic layer was separated and the aqueous layer was extracted
with dichloromethane (15 mL.times.2). The combined organic layer
was washed with saturated brine (50 mL.times.2), dried over
anhydrous sodium sulfate and then filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep-TLC (dichloromethane/methanol=20/1)
gavel-((2-(trimethylsilyl) ethoxy) methyl)-5-(6-(pyridin-2-ylamino)
pyrimidin-4-ylamino)-6-methoxyindazole 20c (6.0 mg, 0.013 mmol,
yellow oil). Yield: 46%.
[0264] MS m/z (ESI): 464 [M+1],
[0265] Step 3
5-(6-(Pyridin-2-ylamino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole
[0266] 1-((2-(trimethylsilyl) ethoxy)
methyl)-5-(6-(pyridin-2-ylamino)
pyrimidin-4-ylamino)-6-methoxyindazole 20c (6.0 mg, 0.013 mmol),
dichloromethane (1 mL) and trifluoroacetic acid (1 mL) were mixed
and stirred at room temperature for 3 hours. Then it was quenched
with saturated aqueous sodium bicarbonate (10 mL). The organic
layer was separated and the aqueous layer was extracted with
dichloromethane (15 mL.times.2). The combined organic layer was
washed with brine (50 mL.times.2), dried over anhydrous sodium
sulfate and then filtered. The filtrate was concentrated under
reduced pressure. Purification of the residue using Prep-HPLC
(water (0.2% formic acid), 10%.about.40% acetonitrile, 15 minutes)
gave 5-(6-(pyridin-2-ylamino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole formate 20 (2.0 mg,
0.006 mmol, white solid). Yield: 46%.
[0267] MS m/z (ESI): 334 [M+1],
[0268] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.83 (s, 1H), 9.65
(s, 1H), 8.42 (s, 1H), 8.20 (s, 1H,), 8.17 (d, J=7.2 Hz, 1H), 8.14
(s, 1H), 7.96 (s, 1H), 7.94 (s, 1H), 7.68-7.64 (m, 1H), 7.52 (d,
J=8.4 Hz, 1H), 7.17 (s, 1H), 7.04 (s, 1H), 6.90-6.85 (m, 1H), 3.87
(s, 3H).
Example 21
5-(6-((4-Chloropyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole
##STR00054##
[0270] Step 1
4-(4-Chloropyridin-2-yl) amino-6-aminopyrimidine
[0271] To the mixture of 4,6-diaminopyrimidine 21a (70.0 mg, 0.6
mmol), 4-chloro-2-fluoropyridine (65.0 mg, 0.5 mmol) and
N,N-dimethylacetamide (5 mL) was added cesium carbonate (327.0 mg,
1 mmol) at room temperature. The reaction was stirred at
110.degree. C. for 3 hours and quenched with water (20 mL). The
organic layer was separated and the aqueous layer was extracted
with dichloromethane (15 mL.times.2). The combined organic layer
was washed with saturated brine (50 mL.times.2), dried over
anhydrous sodium sulfate and then filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep-TLC (dichloromethane/methanol=10/1) gave
4-(4-chloropyridin-2-yl) amino-6-aminopyrimidine 21b (18.0 mg, 0.08
mmol, yellow solid). Yield: 16%.
[0272] MS m/z (ESI): 222 & 224 [M+1],
[0273] Step 2
1-tert-Butoxycarbonyl-5-(6-((4-chloropyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxyindazole
[0274] To the mixture of 4-(4-chloropyridin-2-yl)
amino-6-aminopyrimidine 21b (18.0 mg, 0.08 mmol),
1-tert-butoxycarbonyl-5-bromo-6-methoxyindazole (33.0 mg, 0.1 mmol)
and 1,4-dioxane (2.0 mL) were added
tris(dibenzylideneacetone)-dipalladium(0) (9.0 mg, 0.01 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (11.0 mg, 0.02
mmol) and cesium carbonate (65.0 mg, 0.2 mmol) at room temperature
under argon. The mixture was stirred at 130.degree. C. in microwave
oven under argon for 1 hour and cooled to room temperature, then
quenched with water (10 mL). The organic layer was separated and
the aqueous layer was extracted with dichloromethane (15
mL.times.2). The combined organic layer was washed with saturated
brine (50 mL.times.2), dried over anhydrous sodium sulfate and then
filtered. The filtrate was concentrated under reduced pressure.
Purification of the residue using Prep TLC
(dichloromethane/methanol=20/1) gave
1-tert-butoxycarbonyl-5-(6-((4-chloropyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxyindazole 21c (20 mg, 0.043 mmol,
yellow solid). Yield: 54%.
[0275] MS m/z (ESI): 468 & 470 [M+1],
[0276] Step 3
5-(6-((4-Chloropyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole
[0277] The mixture of
1-tert-butoxycarbonyl-5-(6-((4-chloropyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxyindazole 21c (20.0 mg, 0.043 mmol),
dichloromethane (1.0 mL) and trifluoroacetic acid (1.0 mL) was
stirred for 3 hours at room temperature. Then it was quenched with
saturated aqueous sodium bicarbonate (10 mL). The organic layer was
separated and the aqueous layer was extracted with dichloromethane
(15 mL.times.2). The combined organic layer was washed with
saturated brine (50 mL.times.2), dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated under reduced
pressure. Purification of the residue using Prep TLC
(dichloromethane/methanol=10/1) gave 5-(6-((4-chloropyridin-2-yl)
amino) pyrimidin-4-ylamino)-6-methoxy-1H-indazole 21 (3.0 mg, 0.008
mmol, white solid). Yield: 19%.
[0278] MS m/z (ESI): 368 & 370 [M+1],
[0279] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.86 (s, 1H), 9.91
(s, 1H), 8.56 (s, 1H), 8.27 (s, 1H), 8.18 (d, J=5.2 Hz, 1H), 7.95
(d, J=5.2 Hz, 1H), 7.93 (s, 1H), 7.83 (s, 1H), 7.04 (s, 1H), 7.02
(s, 1H), 6.98 (s, 1H), 3.87 (s, 3H).
Example 22
5-(6-(pyrimidin-4-ylamino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole formate
##STR00055##
[0281] Step 1
1-((2-(trimethylsilyl) ethoxy) methyl)-5-(6-(pyrimidin-4-ylamino)
pyrimidin-4-ylamino)-6-methoxyindazole
[0282] To the mixture of 1-((2-(trimethylsilyl) ethoxy)
methyl)-5-(6-aminopyrimidin-4-ylamino)-6-methoxyindazole 22a (15.0
mg, 0.04 mmol), 4-chloropyrimidine (6.0 mg, 0.04 mmol) and
1,4-dioxane (1.0 mL) were added
tris(dibenzylideneacetone)-dipalladium(0) (3.0 mg, 0.004 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (4.0 mg, 0.008
mmol) and sodium t-butoxide (20.0 mg, 0.2 mmol) at room temperature
under argon. The mixture was reacted at 90.degree. C. in microwave
oven under argon for 1 hour. It was cooled to room temperature and
quenched with water (10 mL). The organic layer was separated and
the aqueous layer was extracted with dichloromethane (15
mL.times.2). The combined organic layer was washed with saturated
brine (50 mL.times.2), dried over anhydrous sodium sulfate and then
filtered, the filtrate was concentrated under reduced pressure.
Purification of the residue using Prep TLC
(dichloromethane/methanol=20/1) gavel-((2-(trimethylsilyl) ethoxy)
methyl)-5-(6-(pyrimidin-4-ylamino)
pyrimidin-4-ylamino)-6-methoxyindazole 22b (10 mg, 0.021 mmol,
yellow solid). Yield: 54%.
[0283] MS m/z (ESI): 465 [M+1],
[0284] Step 2
5-(6-(pyrimidin-4-ylamino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole formate
[0285] 1-((2-(trimethylsilyl) ethoxy)
methyl)-5-(6-(pyrimidin-4-ylamino)
pyrimidin-4-ylamino)-6-methoxyindazole 22b (10.0 mg, 0.021 mmol),
dichloromethane (1 mL) and trifluoroacetic acid (1 mL) were mixed
and stirred at room temperature for 3 hours. The mixture was
quenched with saturated aqueous sodium bicarbonate (10 mL). The
organic layer was separated and the aqueous layer was extracted
with dichloromethane (15 mL.times.2). The combined organic layer
was washed with saturated brine (30 mL), dried over anhydrous
sodium sulfate and filtered. The filtrate was concentrated under
reduced pressure. Purification of the residue using Prep-HPLC
(water (0.2% formic acid), 10%.about.40% acetonitrile, 15 minutes)
gave 5-(6-(pyrimidin-4-ylamino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole formate 22 (3.0 mg,
0.009 mmol, white solid). Yield: 46%.
[0286] MS m/z (ESI): 335 [M+1],
[0287] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.99 (s, 1H), 10.15
(s, 1H), 8.67 (s, 2H), 8.44 (d, J=6.0 Hz, 1H), 8.29 (s, 1H), 8.17
(s, 1H), 7.96 (s, 1H), 7.94 (s, 1H), 7.66 (d, J=6.0 Hz, 1H), 7.08
(s, 1H), 7.05 (s, 1H), 3.87 (s, 3H).
Example 23
5-(6-((6-Chloropyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole
##STR00056##
[0289] The synthetic procedure is similar to that in Example 22.
using 2,4-dichloropyridine instead of 4-chloropyrimidine, the
desired compound 23: 5-(6-((6-chloropyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole (4.0 mg, 0.011 mmol,
white solid) was obtained. Yield: 79%.
[0290] MS m/z (ESI): 368 [M+1],
[0291] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.79 (s, 1H), 9.91
(s, 1H), 8.53 (s, 1H), 8.24 (s, 1H), 7.94 (s, 1H), 7.82 (s, 1H),
7.70-7.68 (m, 1H), 7.58 (d, J=8.2 Hz, 1H), 7.06 (s, 1H), 6.94 (d,
J=7.5 Hz, 1H), 6.91 (s, 1H), 3.86 (s, 3H).
Example 24
5-(6-((5-Chloropyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole formate
##STR00057##
[0293] Step 1
N-(5-Chloropyridin-2-yl) pyrimidine-4,6-diamine
[0294] To the mixture of 4,6-diaminopyrimidine 24a (110.0 mg, 1.0
mmol), sodium hydride (60% dispersion in mineral oil, 120.0 mg, 5.0
mmol) and N,N-dimethylacetamide (10.0 mL) was added
2,5-dichloropyridine (6.0 mg, 0.03 mmol) at room temperature. The
mixture was reacted at 70.degree. C. for 2 hours. Then it was
cooled to room temperature and quenched with saturated aqueous
sodium bicarbonate (10 mL). The mixture was extracted with
dichloromethane (30 mL.times.3). The combined organic layer was
washed with saturated brine (30 mL), dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated under reduced
pressure. Purification of the residue using flash column
chromatography (dichloromethane/methanol=10/1) gave
N-(5-chloropyridin-2-yl) pyrimidine-4,6-diamine 24b (45.0 mg, 0.20
mmol, white solid). Yield: 20%.
[0295] MS m/z (ESI): 222 & 224 [M+1],
[0296] Step 2
1-tert-Butoxycarbonyl-5-(6-((5-chloropyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxyindazole
[0297] To the mixture of N-(5-chloropyridin-2-yl)
pyrimidine-4,6-diamine 24b (22.0 mg, 0.10 mmol),
1-tert-butoxycarbonyl-5-bromo-6-methoxyindazole (32.0 mg, 0.10
mmol) and 1,4-dioxane (2.0 mL) were added
tris(dibenzylideneacetone)-dipalladium(0) (9.0 mg, 0.01 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (12.0 mg, 0.02
mmol) and cesium carbonate (98.0 mg, 0.3 mmol) under argon. The
reaction mixture was reacted at 110.degree. C. in microwave oven
under argon for 1 hour. Then it was diluted with dichloromethane
(10 mL) and filtered. The filtrate was concentrated under reduced
pressure. Purification of the residue using Prep TLC
(dichloromethane/methanol=20/1)
gavel-tert-butoxycarbonyl-5-(6-((5-chloropyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxyindazole 24c (10 mg, 0.02 mmol, white
solid). Yield: 20%.
[0298] MS m/z (ESI): 468 & 470 [M+1],
[0299] Step 3
5-(6-((5-Chloropyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole formate
[0300] 1-tert-Butoxycarbonyl-5-(6-((5-chloropyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxyindazole 24c (10.0 mg, 0.02 mmol),
dichloromethane (2 mL) and trifluoroacetic acid (1 mL) were mixed
and stirred at room temperature for 3 hours. Then the mixture was
quenched with saturated aqueous sodium bicarbonate (5 mL). The
organic layer was separated and the aqueous layer was extracted
with dichloromethane (10 mL.times.2). The combined organic layer
was washed with saturated brine (10 mL), dried over anhydrous
sodium sulfate and filtered. The filtrate was concentrated under
reduced pressure. Purification of the residue using Prep-HPLC
(water (0.2% formic acid), 10%.about.40% acetonitrile, 15 minutes)
gave 5-(6-((5-chloropyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole formate 24 (5.0 mg,
0.014 mmol, white solid). Yield: 70%.
[0301] MS m/z (ESI): 368 & 370 [M+1],
[0302] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.85 (s, 1H), 9.85
(s, 1H), 8.50 (s, 1H), 8.24 (s, 2H), 8.19 (s, 1H), 7.95 (s, 1H),
7.94 (s, 1H), 7.77 (s, 1H), 7.68 (s, 1H), 7.04 (d, J=9.2 Hz, 1H),
6.98 (d, J=9.2 Hz, 1H), 3.87 (s, 3H).
Example 25
5-(6-((4-(Trifluoromethyl) pyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole
##STR00058##
[0304] Step 1
1-((2-(Trimethylsilyl) ethoxy) methyl)-5-(6-((4-(trifluoromethyl)
pyridin-2-yl) amino) pyrimidin-4-yl Amino)-6-methoxyindazole
[0305] To the mixture of 1-((2-(trimethylsilyl) ethoxy)
methyl)-5-(6-aminopyrimidin-4-ylamino)-6-methoxyindazole 25a (15.0
mg, 0.039 mmol), 2-chloro-4-(trifluoromethyl)pyridine (10.7 mg,
0.059 mmol) and 1,4-dioxane (1 mL) were added
tris(dibenzylideneacetone)dipalladium(0) (3.6 mg, 0.004 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (2.3 mg, 0.004
mmol) and sodium t-butoxide (7.5 mg, 0.078 mmol) under argon. The
reaction mixture was stirred at 80.degree. C. in microwave oven
under argon for 1 hour. Then it was cooled to room temperature,
diluted with dichloromethane (10 mL) and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep-HPLC (water (0.2% formic acid), 40%-60% acetonitrile, 15
minutes) gave 1-((2-(trimethylsilyl) ethoxy)
methyl)-5-(6-((4-(trifluoromethyl) pyridin-2-yl) amino)
pyrimidin-4-yl Amino)-6-methoxyindazole 25b (5.5 mg, 0.01 mmol,
white solid).
[0306] Yield: 26%.
[0307] MS m/z (ESI): 532 [M+1],
[0308] Step 2
5-(6-((4-(Trifluoromethyl) pyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole
[0309] 1-((2-(Trimethylsilyl) ethoxy)
methyl)-5-(6-((4-(trifluoromethyl) pyridin-2-yl) amino)
pyrimidin-4-yl Amino)-6-methoxyindazole 25b (5.5 mg, 0.010 mmol)
was dissolved in dichloromethane (1 mL) and trifluoroacetic acid (1
mL) was added. The mixture was stirred at room temperature for 2
hours. Then it was concentrated under reduced pressure. The residue
was dissolved in dimethyl sulfoxide (1 mL) and adjusted to
pH=8.about.9 with one drop of saturated sodium hydroxide solution.
Purification of the mixture using Prep-HPLC (water (0.2% formic
acid), 30%.about.70% acetonitrile, 15 minutes) gave
5-(6-((4-(trifluoromethyl) pyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole 25 (1.6 mg, 0.004 mmol,
white solid). Yield: 40%.
[0310] MS m/z (ESI): 402 [M+1],
[0311] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.85 (s, 1H), 10.09
(s, 1H), 8.58 (s, 1H), 8.43 (d, J=5.2 Hz, 1H), 8.28 (s, 1H), 8.06
(s, 1H), 8.02-7.88 (m, 2H), 7.20 (d, J=5.2 Hz, 1H), 7.04 (s, 1H),
7.00 (s, 1H), 3.87 (s, 3H).
Example 26
5-(6-((5-chloropyrimidin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole
##STR00059##
[0313] The synthetic procedure is similar to that of example 22.
Using 2,5-dichloropyrimidine instead of 2, 5-dichloropyridine and
the desired compound 26: 5-(6-((5-chloropyrimidin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole (4.0 mg, 0.01 mmol,
white solid) was obtained. Yield: 50%.
[0314] MS m/z (ESI): 369 & 371 [M+1],
[0315] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.85 (s, 1H), 10.29
(s, 1H), 8.80 (s, 1H), 8.64 (s, 2H), 8.28 (s, 1H), 8.00 (s, 1H),
7.96 (s, 1H), 7.47 (s, 1H), 7.05 (s, 1H), 3.87 (s, 3H).
Example 27
5-(6-(Pyrimidin-2-ylamino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole
##STR00060##
[0317] The synthetic procedure is similar to that of example 20.
Using 2-chloropyrimidine instead of 2-bromopyridine, and the
desired compound 5-(6-(pyrimidin-2-ylamino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole 27 was obtained. Yield:
63%.
[0318] MS m/z (ESI): 335 [M+1],
[0319] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 13.03 (s, 1H), 11.25
(s, 1H), 10.07 (s, 1H), 8.70 (s, 1H), 8.69 (s, 1H), 8.51 (s, 1H),
8.02 (s, 1H), 7.82 (s, 1H), 7.23 (t, J=5.2 Hz, 1H), 7.13 (s, 1H),
7.04 (s, 1H), 3.88 (s, 3H). Example 28
5-(6-((3-Chloropyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole
##STR00061##
[0321] The synthetic procedure is similar to that of example 24.
Using 2,3-dichloropyridine instead of 2,5-dichloropyridine, and the
desired compound: 5-(6-((3-chloropyridin-2-yl) amino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole 28 was obtained. Yield:
76%.
[0322] MS m/z (ESI): 368 & 370 [M+1],
[0323] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.85 (s, 1H), 8.65
(s, 1H), 8.40 (s, 1H), 8.30 (s, 1H), 8.22 (d, J=6.4 Hz, 1H), 8.01
(s, 1H), 7.93 (d, J=10 Hz, 1H), 7.38 (s, 1H), 7.09-7.05 (m, 1H),
7.03 (s, 1H), 6.68 (s, 1H), 3.87 (s, 3H).
Example 29
5-(6-(Pyrazin-2-ylamino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole
##STR00062##
[0325] The synthetic procedure is similar to that of example 20.
Using 2-chloropyrazine instead of 2-chloropyridine, and the desired
compound: 5-(6-(pyrazin-2-ylamino)
pyrimidin-4-ylamino)-6-methoxy-1H-indazole 29 was obtained. Yield:
31%.
[0326] MS m/z (ESI): 335 [M+1],
[0327] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.86 (s, 1H), 10.02
(s, 1H), 8.90 (s, 1H), 8.61 (s, 1H), 8.31 (s, 1H), 8.27 (s, 1H),
8.20 (d, J=2.4 Hz, 1H), 8.10 (d, J=2.4 Hz, 1H), 7.95 (s, 1H), 7.07
(s, 1H), 7.04 (s, 1H), 3.87 (s, 3H).
Example 30
5-(6-Methylaminopyrimidin-4-ylamino)-6-methoxy-1H-indazole
##STR00063##
[0329] Step 1
1-tert-Butoxycarbonyl-5-(6-chloropyrimidin-4-ylamino)-6-methoxyindazole
[0330] 4,6-Dichloropyrimidine 30a (26.3 mg, 0.1 mmol),
1-tert-butoxycarbonyl-5-amino-6-methoxyindazole (29.8 mg, 0.2
mmol), cesium carbonate (97.8 mg, 0.3 mmol) and dimethyl acetamide
(1 mL) were mixed and stirred at 100.degree. C. in microwave oven
for 1 hour. Then it was diluted water (10 mL) and extracted with
dichloromethane (10 mL.times.3). The combined organic layer was
washed with saturated brine (10 mL.times.2), dried over anhydrous
sodium sulfate and filtered. The filtrate was concentrated under
reduced pressure. Purification of the residue using Prep-TLC
(dichloromethane/methanol=20/1)
gavel-tert-butoxycarbonyl-5-(6-chloropyrimidin-4-ylamino)-6-methoxyindazo-
le 30b (10 mg, 0.027 mmol, yellow solid). Yield: 27%.
[0331] MS m/z (ESI): 376 & 378 [M+1],
[0332] Step 2
5-(6-Methylaminopyrimidin-4-ylamino)-6-methoxy-1H-indazole
[0333]
1-tert-Butoxycarbonyl-5-(6-chloropyrimidin-4-ylamino)-6-methoxyinda-
zole 30b (10 mg, 0.027 mmol) and methanamine (2 mL, 8 mmol, 4 M
solution in ethanol) were mixed and stirred at 70.degree. C. for 5
hours. Then it was concentrated under reduced pressure.
Purification of the residue using Prep-HPLC (water (0.2% formic
acid), 20%.about.70% acetonitrile, 15 minutes) gave
5-(6-methylamino-pyrimidin-4-ylamino)-6-methoxy-1H-indazole 30 (4
mg, 0.015 mmol, white solid).
[0334] Yield: 56%.
[0335] MS m/z (ESI): 271 [M+1],
[0336] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.82 (s, 1H),
8.02-7.91 (m, 4H), 7.00 (s, 1H), 6.67-6.61 (m, 1H), 5.61 (s, 1H),
3.87 (s, 3H), 2.68-2.67 (d, J=4.4 Hz, 3H).
Example 31
5-(6-Cyclopropylaminopyrimidin-4-ylamino)-6-methoxy-1H-indazole
formate
##STR00064##
[0338] The synthetic procedure is similar to that of example 30.
Using cyclopropylamine instead of methylamine, and the desired
compound
5-(6-cyclopropylamino-pyrimidin-4-ylamino)-6-methoxy-1H-indazole
formate 31 was obtained. Yield: 37%, Prep HPLC conditions (water
(0.2% formic acid), 20% to 70% acetonitrile, 15 minutes).
[0339] MS m/z (ESI): 297 [M+1],
[0340] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.80 (s, 1H), 8.09
(s, 1H), 8.05 (s, 1H), 8.00 (s, 1H), 7.91 (s, 1H), 7.01 (s, 1H),
6.97 (s, 1H), 6.07 (s, 1H), 5.89 (s, 1H), 3.88 (s, 3H), 2.40-2.31
(m, 1H), 0.72-0.56 (m, 2H), 0.44-0.38 (m, 2H).
Example 32
5-((6-Amino-5-cyclohex-3-ene-1-carboxypyrimidin-4-yl)
amino)-6-methoxy-1H-indazole hydrochloride
##STR00065##
[0342] Step 1
Ethyl
4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate
[0343] To the mixture of pyridine (1.7 g, 21.18 mmol) in anhydrous
toluene (15.0 mL) was added dropwise trifluoromethanesulfonic
anhydride (5.9 g, 21.28 mmol) in an ice bath. The mixture was
stirred at room temperature for 30 minutes, then the solution of
ethyl 4-oxocyclohexane-1-carboxylate 32a (3.0 g, 17.65 mmol) in
anhydrous toluene (15 mL) was added to the mixture. It was stirred
at 50.degree. C. for 12 hours. Then it was quenched with water (30
mL). The organic layer was separated. Silica gel (15 g) was added
to the organic layer and filtered. The filter cake was washed with
toluene (10 mL.times.3). The combined organic layer was dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated under reduced pressure to give ethyl
4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate 32b
(3 g, 9.93 mmol, yellow oil). Yield: 56%.
[0344] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.79-5.77 (m, 1H),
4.16 (q, J=7.1 Hz, 2H), 2.64-2.53 (m, 1H), 2.48-2.38 (m, 4H),
2.18-2.09 (m, 1H), 1.99-1.86 (m, 1H), 1.27 (t, J=7.1 Hz, 3H).
[0345] Step 2
1-Ethoxycarbonylcyclohex-3-ene-4-boronic acid pinacol ester
[0346] To the mixture of ethyl
4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate 32b
(1.0 g, 3.31 mmol), bis(pinacolato)diboron (1.2 g, 5.30 mmol) and
potassium acetate (0.7 g, 6.62 mmol) in 1,4-dioxane (10.0 mL) was
added [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II)
(0.24 g, 0.16 mmol) under agon. The reaction mixture was stirred at
90.degree. C. for 12 hours. Then it was cooled to room temperature
and filtered. The filtrate was concentrated under reduced pressure.
Purification of the residue using Prep-TLC (n-hexane/ethyl
acetate=10/1) gave 1-ethoxycarbonylcyclohex-3-ene-4-boronic acid
pinacol ester 32c (0.20 g, 0.71 mmol, colorless oil). Yield:
21%.
[0347] MS m/z (ESI): 281 [M+1],
[0348] Step 3
4,6-Diamino-5-iodopyrimidine
[0349] To the mixture of 4,6-diaminopyrimidine 32d (1.0 g, 9.10
mmol) and potassium carbonate (1.9 g, 13.70 mmol) in water (20.0
mL) and N,N-dimethylformamide (10.0 mL) was added iodine (2.6 g,
10.00 mmol) at room temperature. The mixture was stirred at
45.degree. C. for 18 hours. Then it was quenched with saturated
aqueous sodium bisulfite (10 mL) and filtered. The filter cake was
washed with water (40 mL) to give 4,6-diamino-5-iodopyrimidine 32e
(1.2 g, 5.08 mmol, white solid). Yield: 56%.
[0350] MS m/z (ESI): 237 [M+1],
[0351] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.72 (s, 1H),
6.35-6.25 (brs, 4H).
[0352] Step 4
Ethyl 4-(4,6-diaminopyrimidin-5-yl)cyclohex-3-ene-1-carboxylate
[0353] To the solution of 4,6-diamino-5-iodopyrimidine 32e (80.0
mg, 0.34 mmol), 1-ethoxycarbonylcyclohex-3-ene-4-boronic acid
pinacol ester 32c (120.0 mg, 0.44 mmol) and cesium carbonate (320.0
mg, 0.10 mmol) in 1,4-dioxane (5.0 mL) and water (1.0 mL) were
added palladium(II) acetate (8.0 mg, 0.03 mmol) and
2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (28.0 mg, 0.06 mol)
under argon. The mixture was stirred at 50.degree. C. under argon
for 12 hours. Then it was cooled to room temperature and filtered.
The filtrate was concentrated under reduced pressure. Purification
of the residue using Prep-TLC (dichloromethane/methanol=15/1) gave
ethyl 4-(4,6-diaminopyrimidin-5-yl)cyclohex-3-ene-1-carboxylate 32f
(15 mg, 0.19 mmol, yellow solid). Yield: 56%.
[0354] MS m/z (ESI): 263 [M+1],
[0355] Step 5
5-((6-Amino-5-cyclohex-3-ene-1-carboxypyrimidin-4-yl)
amino)-6-methoxy-1H-indazole hydrochloride
[0356] To the solution of ethyl
4-(4,6-diaminopyrimidin-5-yl)cyclohex-3-ene-1-carboxylate 32f (20
mg, 0.08 mmol),
1-tert-butoxycarbonyl-5-((6-acetylaminopyrimidin-4-yl)
amino)-6-methoxy-1H-indazole (25 mg, 0.08 mmol) and sodium
t-butoxide (30 mg, 0.30 mmol) in 1,4-dioxane (1.0 mL) were added
tris(dibenzylideneacetone)dipalladium(0) (7 mg, 0.008 mmol) and
2-(dicyclohexylphosphine) 3,6-dimethoxy-2', 4',
6'-triisopropyl-1,1'-biphenyl(9 mg, 0.015 mmol) under argon. The
mixture was stirred at 100.degree. C. under argon for 12 hours.
Then it was cooled to room temperature and adjusted to pH=5.about.6
with diluted hydrochloric acid (1 M). The mixture was concentrated
under reduced pressure. Purification of the residue using Prep-HPLC
(water (0.2% formic acid), 20%-40% acetonitrile, 15 minutes) gave
5-((6-amino-5-cyclohex-3-ene-1-carboxypyrimidin-4-yl)
amino)-6-methoxy-1H-indazole hydrochloride 32 (2 mg, 0.005 mmol,
white solid). Yield: 6%.
[0357] MS m/z (ESI): 381 [M+1],
Example 33
5-((6-Amino-5-cyclohex-3-en-1-yl (4-methylpiperazin-1-yl)
methanylpyrimidin-4-yl) amino)-6-methoxy-1H-indazole formate
##STR00066##
[0359] 5-((6-Amino-5-cyclohex-3-ene-1-carboxypyrimidin-4-yl)
amino)-6-methoxy-1H-indazole hydrochloride 32 (7.0 mg, 0.02 mmol),
2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (16.0 mg, 0.04 mmol), N,N-diisopropylethylamine
(13.0 mg, 0.1 mmol) and N,N-dimethylformamide (1.0 mL) were mixed
and stirred at room temperature for 10 minutes. Then
N-methylpiperazine (20.0 mg, 0.2 mmol) was added to the above
mixture and stirred at room temperature for 24 hours. The mixture
was diluted with water (10 mL) and extracted with dichloromethane
(10 mL.times.3). The combined organic layer was washed with
saturated brine (10 mL), dried over anhydrous sodium sulfate and
filtered. The filtrate was concentrated under reduced pressure.
Purification of the residue using Prep-HPLC (water (0.2% formic
acid), 10%.about.40% acetonitrile, 15 minutes) gave
5-((6-amino-5-cyclohex-3-en-1-yl (4-methylpiperazin-1-yl)
methanylpyrimidin-4-yl) amino)-6-methoxy-1H-indazole formate 33
(2.0 mg, 0.005 mmol, white solid). Yield: 25%.
[0360] MS m/z (ESI): 463 [M+1],
[0361] .sup.1H NMR (400 MHz, CD.sub.3OD) b 8.39 (s, 1H), 8.25 (s,
1H), 8.05 (s, 1H), 8.01 (s, 1H), 7.95 (s, 1H), 7.07 (s, 1H), 6.02
(s, 1H), 3.96 (s, 3H), 3.87-3.80 (m, 4H), 3.65-3.60 (m, 1H),
3.24-3.16 (m, 2H), 2.95-2.88 (m, 4H), 2.65 (s, 3H), 2.52-3.40 (m,
3H), 2.24-2.19 (m, 1H).
Example 34
4-(4-Amino-6-((6-methoxy-1H-indazol-5-yl)amino)pyrimidin-5-yl)-N,N-dimethy-
lcyclohex-3-ene-1-carboxamide formate
##STR00067##
[0363] 5-((6-Amino-5-cyclohex-3-ene-1-carboxypyrimidin-4-yl)
amino)-6-methoxy-1H-indazole hydrochloride 32 (7.0 mg, 0.02 mmol),
2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (11.0 mg, 0.03 mmol), dimethylamine
hydrochloride (5.0 mg, 0.06 mmol), diisopropylethylamine (10.0 mg,
0.07 mmol) were dissolved into N,N-dimethylformamide (1.0 mL) and
stirred at room temperature for 12 hours. Then it was concentrated
under reduced pressure. Purification of the residue using Prep-HPLC
(water (0.2% formic acid), 20%-50% acetonitrile, 15 minutes) gave
4-(4-amino-6-((6-methoxy-1H-indazol-5-yl)amino)pyrimidin-5-yl)-N,N-dimeth-
ylcyclohex-3-ene-1-carboxamide formate 34 (2.5 mg, 0.006 mmol white
solid). Yield: 31%.
[0364] MS m/z (ESI): 408 [M+1],
[0365] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.65 (brs, 2H), 8.59
(s, 1H), 8.07 (s, 1H), 8.01 (s, 1H), 7.85 (s, 1H), 7.42-7.32 (m,
1H), 6.93 (s, 1H), 6.03-5.95 (m, 2H), 3.86 (s, 3H), 3.05 (s, 3H),
2.80 (s, 3H), 2.32-2.04 (m, 4H), 1.94-1.62 (m, 3H).
Example 35
1-tert-Butoxycarbonyl-5-((6-amino-5-ethoxycarbonylpyrimidin-4-yl)
amino)-6-methoxyindazole
##STR00068##
[0367] Step 1
4-Aminopyrimidine-5-carbonitrile
[0368] To the mixture of malononitrile 35a (20.8 g, 200 mmol),
formamidine acetate (6.6 g, 100 mmol) and methanol (400 mL) was
added in portions sodium methanolate (13.5 g, 250 mmol) at room
temperature. The mixture was stirred at room temperature for 48
hours. Then it was filtered. The filter cake was washed with ice
water (200 mL) and dried to give 4-aminopyrimidine-5-carbonitrile
35b (5.4 g, 45 mmol, yellow solid). Yield: 45%.
[0369] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.59 (s, 1H), 8.53
(s, 1H), 7.91 (s, 2H).
[0370] Step 2
Pyrimido[4,5-d]pyrimidin-4-amine
[0371] 4-aminopyrimidine-5-carbonitrile 35b (2.4 g, 20 mmol),
formamidine acetate (3.1 g, 30 mmol) and ethylene glycol (20 mL)
were mixed and stirred at 140.degree. C. under argon for 0.5 hour.
Then it was filtered. The filter cake was washed with methanol (50
mL) and dried to give pyrimido[4,5-d]pyrimidin-4-amine 35c (1.5 g,
10 mmol, yellow solid). Yield: 50%.
[0372] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.71 (s, 1H), 9.34
(s, 1H), 8.71-8.49 (m, 3H).
[0373] Step 3
4,6-Diaminopyrimidine-5-carbaldehyde
[0374] Pyrimido[4,5-d]pyrimidin-4-amine 35c (1.0 g, 7 mmol) and
hydrochloric acid aqueous solution (20 mL, 0.5 M) were mixed at
room temperature and stirred at 100.degree. C. for 2 hours. Then it
was quenched with 1 M aqueous sodium hydroxide. The solid formed
was filtered. The filter cake was washed with water (50 mL) and
dried to give 4,6-diaminopyrimidine-5-carbaldehyde 35d (450.0 mg,
3.4 mmol, yellow solid). Yield: 48%.
[0375] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.06 (s, 1H), 7.90
(s, 1H), 7.74 (s, 4H).
[0376] Step 4
4,6-Diaminopyrimidine-5-carboxylic acid
[0377] To the mixture of 4,6-diaminopyrimidine-5-carbaldehyde 35d
(0.4 g, 3.0 mmol) and ammonium chlorite (0.14 g, 15 mmol) in
dichloromethane (20 mL) and water (20 mL) was added dropwise
phosphoric acid (1.5 mL) and dimethyl sulfoxide (3 mL) at room
temperature. The mixture was stirred at room temperature for 24
hours. Then it was quenched with saturated aqueous sodium
bicarbonate (50 mL) and filtered. The filter cake was washed with
water (50 mL) and dried to give 4,6-diaminopyrimidine-5-carboxylic
acid 35e (0.3 g, 1.8 mmol, yellow solid). Yield: 60%.
[0378] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 13.14 (s, 1H), 8.70
(s, 4H), 8.26 (s, 1H).
[0379] Step 5
Ethyl 4,6-diaminopyrimidine-5-carboxylate
[0380] To the mixture of 4,6-diaminopyrimidine-5-carboxylic acid
35e (462.0 mg, 3 mmol) in dichloromethane (20 mL) was added slowly
N,N-dimethylformamide (5 d) and oxalyl chloride (1.5 mL) at room
temperature. The mixture was stirred at room temperature for 0.5
hour. Then anhydrous ethanol was added and stirred at room
temperature for 0.5 hour. Then it was quenched with saturated
aqueous sodium dicarbonate (50 mL). The organic layer was
separated, and the aqueous layer was extracted with dichloromethane
(50 mL.times.3). The combined organic layer was washed with
saturated brine (50 mL), dried with anhydrous sodium sulfate and
filtered. The filtrate was concentrated under reduced pressure.
Purification of the residue using flash column chromatography
(dichloromethane/methanol=20:1) gave ethyl
4,6-diaminopyrimidine-5-carboxylate 35f (280.0 mg, 1.5 mmol, white
solid). Yield: 51%.
[0381] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.02 (s, 1H), 7.66
(s, 2H), 5.68 (s, 2H), 4.39 (q, J=7.2 Hz, 2H), 1.42 (t, J=7.2 Hz,
3H).
[0382] Step 6
1-tert-Butoxycarbonyl-5-((6-amino-5-ethoxycarbonylpyrimidin-4-yl)
amino)-6-methoxyindazole
[0383] To the mixture of ethyl 4,6-diaminopyrimidine-5-carboxylate
35f (18.0 mg, 0.1 mmol),
1-tert-butoxycarbonyl-5-bromo-6-methoxyindazole (232.0 mg, 0.10
mmol) and 1,4-dioxane (2.0 mL) were added
tris(dibenzylideneacetone)dipalladium(0) (9 mg, 0.01 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (12 mg, 0.02 mmol)
and cesium carbonate (98.0 mg, 0.30 mmol) under argon. The reaction
mixture was stirred at 110.degree. C. in microwave oven under argon
for 1 hour. Then it was diluted with dichloromethane (10 mL) and
filtered. The filtrate was concentrated under reduced pressure.
Purification of the residue using Prep-TLC (hexane/ethyl
acetate=1/1) gave
1-tert-butoxycarbonyl-5-((6-amino-5-ethoxycarbonylpyrimidin-4-yl)
amino)-6-methoxyindazole 35 (5 mg, 0.01 mmol, white solid). Yield:
10%.
[0384] MS m/z (ESI): 429 [M+1],
[0385] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.76 (s, 1H),
8.94 (s, 1H), 8.24 (s, 1H), 8.10 (s, 1H), 7.75 (s, 1H), 4.50 (q,
J=7.2 Hz, 2H), 4.07 (s, 3H), 1.73 (s, 9H), 1.49 (t, J=7.2 Hz,
3H).
Example 36
5-((6-Amino-5-ethoxycarbonylpyrimidin-4-yl)
amino)-6-methoxy-1H-indazole dicarboxylate
##STR00069##
[0387]
1-tert-Butoxycarbonyl-5-((6-amino-5-ethoxycarbonylpyrimidin-4-yl)
amino)-6-methoxyindazole 35 (4.0 mg, 0.01 mmol), dichloromethane (2
mL) and trifluoroacetic acid (1 mL) were mixed and stirred at room
temperature for 3 hours. Then it was quenched with saturated
aqueous sodium bicarbonate (5 mL) and diluted with dichloromethane
(5 mL). The organic layer was separated and the aqueous layer was
extracted with dichloromethane (10 mL.times.3). The combined
organic layer was washed with saturated brine (10 mL), dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated. Purification of the residue using Prep-HPLC (water
(0.2% formic acid), 30%.about.70% acetonitrile, 15 minutes) gave
5-((6-amino-5-ethoxycarbonylpyrimidin-4-yl)
amino)-6-methoxy-1H-indazole dicarboxylate 36 (1.3 mg, 0.004 mmol,
white solid).
[0388] Yield: 40%.
[0389] MS m/z (ESI): 329 [M+1],
[0390] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.74 (s, 1H), 10.69
(s, 1H), 8.72 (s, 1H), 8.09 (s, 1H), 7.88 (s, 1H), 7.53 (s, 2H),
7.13 (s, 1H), 6.98 (s, 1H), 6.59 (s, 1H), 4.35 (q, J=7.2 Hz, 2H),
3.90 (s, 3H), 1.31 (t, J=7.2 Hz, 3H).
Example 37
1-tert-Butoxycarbonyl-5-((6-(acetylamino)-5-ethoxycarbonylpyrimidin-4-yl)
amino)-6-methoxyindazole
##STR00070##
[0392] Step 1
Ethyl 4-acetamido-6-aminopyrimidine-5-carboxylate
[0393] Ethyl 4,6-diaminopyrimidine-5-carboxylate 37a (128.0 mg, 0.7
mmol), acetic anhydride (72.0 mg, 0.7 mmol) and dioxane (10 mL)
were mixed and stirred at 80.degree. C. for 6 hours. Then it was
cooled to room temperature and concentrated under reduced pressure.
The mixture was extracted with dichloromethane (20 mL.times.3). The
combined organic layer was washed with saturated brine (20 mL),
dried with anhydrous sodium sulfate and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep-TLC (dichloromethane/methanol=20:1) gave ethyl
4-acetamido-6-aminopyrimidine-5-carboxylate 37b (60.0 mg, 0.27
mmol, white solid). Yield: 38%.
[0394] MS m/z (ESI): 225 [M+1],
[0395] Step 2
1-tert-Butoxycarbonyl-5-((6-(acetylamino)-5-ethoxycarbonylpyrimidin-4-yl)
amino)-6-methoxyindazole
[0396] To the mixture of ethyl
4-acetamido-6-aminopyrimidine-5-carboxylate 37b (23.0 mg, 0.1
mmol), 1-tert-butoxycarbonyl-5-bromo-6-methoxyindazole (32.0 mg,
0.10 mmol) and 1,4-dioxane (2.0 mL) were added
tris(dibenzylideneacetone)-dipalladium(0) (9 mg, 0.01 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (12 mg, 0.02 mmol)
and cesium carbonate (98.0 mg, 0.30 mmol) under argon. The mixture
was reacted at 110.degree. C. in microwave oven under argon for 1
hour. Then it was diluted with dichloromethane (10 mL) and
filtered. The filtrate was concentrated under reduced pressure.
Purification of the residue using Prep-TLC (hexane/ethyl
acetate=1:1) gave
1-tert-butoxycarbonyl-5-((6-(acetylamino)-5-ethoxycarbonylpyrimidin-4-yl)
amino)-6-methoxyindazole 37 (6 mg, 0.015 mmol, white solid). Yield:
15%.
[0397] MS m/z (ESI): 471 [M+1],
[0398] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.60 (s, 1H),
10.56 (s, 1H), 8.90 (s, 1H), 8.56 (s, 1H), 8.12 (s, 1H), 7.78 (s,
1H), 4.55 (q, J=7.2 Hz, 2H), 4.07 (s, 3H), 2.51 (s, 3H), 1.74 (s,
9H), 1.52 (t, J=7.2 Hz, 3H).
Example 38
5-((6-(Acetylamino)-5-ethoxycarbonylpyrimidin-4-yl)
amino)-6-methoxy-1H-indazole
##STR00071##
[0400] The synthetic procedure is similar to that of example 2.
Using
1-t-butoxyacyl-5-((6-(acetylamino)-5-ethoxycarbonylpyrimidine-4-yl)amino)-
-6-methoxyindazole instead of
1-t-butoxyacyl-5-(6-acetamidopyrimidine-4-ylamino)-6-methoxyindazole,
the desired compound:
5-((6-(acetylamino)-5-ethoxycarbonylpyrimidin-4-yl)
amino)-6-methoxy-1H-indazole 38 (1.5 mg, 0.004 mmol, white solid)
was obtained. Yield: 40%.
[0401] MS m/z (ESI): 371 [M+1],
[0402] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.57 (s, 1H),
10.45 (s, 1H), 8.76 (s, 1H), 8.54 (s, 1H), 8.03 (s, 1H), 6.96 (s,
1H), 4.54 (q, J=7.2 Hz, 2H), 4.00 (s, 3H), 2.51 (s, 3H), 1.52 (t,
J=7.2 Hz, 3H).
Example 39
5-((6-Amino-5-carbonylpyrimidin-4-yl) amino)-6-methoxy-1H-indazole
formate
##STR00072##
[0404]
1-tert-Butoxycarbonyl-5-((6-amino-5-ethoxycarbonylpyrimidin-4-yl)
amino)-6-methoxyindazole 35 (20.0 mg, 0.05 mmol), lithium hydroxide
(1 mL, 1 M in water) and ethanol (5 mL) were mixed and stirred at
room temperature for 1 hour. Then it was concentrated under reduced
pressure. The residue was diluted with dichloromethane. The mixture
was adjusted to pH=5.about.6 with 0.5 M hydrochloric acid and
extracted with dichloromethane (20 mL.times.3). The combined
organic layer was washed with saturated brine (20 mL), dried with
anhydrous sodium sulfate and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep-HPLC (water (0.2% formic acid), 10%.about.40%
acetonitrile, 15 minutes) gave
5-((6-amino-5-carbonylpyrimidin-4-yl) amino)-6-methoxy-1H-indazole
formate 39 (6.0 mg, 0.02 mmol, white solid). Yield: 40%.
[0405] MS m/z (ESI): 301 [M+1],
[0406] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.71 (s, 1H), 8.63
(s, 1H), 8.37 (s, 1H), 7.29 (s, 1H), 4.10 (s, 3H).
Example 40
5-((6-Amino-5-dimethylcarbamoyl) pyrimidin-4-yl)
amino)-6-methoxy-1H-indazole
##STR00073##
[0408] The synthetic procedure is similar to that of example 34.
Using
4-amino-6-[(6-methoxy-1H-indazol-5-yl)amino]pyrimidine-5-methanoic
acid instead of
4-(4-amino-6-((6-methoxy-1H-indazol-5-yl)amino)pyrimidine-5-yl)cyclohex-3-
-ene-1-methanoic acid. The desired compound:
5-((6-amino-5-dimethylcarbamoyl) pyrimidin-4-yl)
amino)-6-methoxy-1H-indazole 40 (1.7 mg, 0.005 mmol, white solid)
was obtained. Yield: 25%.
[0409] MS m/z (ESI): 328 [M+1],
[0410] .sup.1H NMR (400 MHz, CD.sub.3OD) b 8.09 (s, 1H), 8.00 (s,
1H), 7.98 (s, 1H), 7.19 (s, 1H), 3.95 (s, 3H), 3.10 (s, 6H).
Example 41
5-((6-Amino-5-(4-morpholino) keto) pyrimidin-4-yl)
amino)-6-methoxy-1H-indazole
##STR00074##
[0412] The synthetic procedure is similar to that of example 34.
Using
4-amino-6-[(6-methoxy-1H-indazol-5-yl)amino]pyrimidine-5-methanoic
acid instead of
4-(4-amino-6-((6-methoxy-1H-indazol-5-yl)amino)pyrimidine-5-yl)cyclohex-3-
-ene-1-methanoic acid. The desired compound:
5-((6-amino-5-(4-morpholino) keto) pyrimidin-4-yl)
amino)-6-methoxy-1H-indazole 41 (1.5 mg, 0.004 mmol, white solid)
was obtained. Yield: 20%.
[0413] MS m/z (ESI): 370 [M+1];
[0414] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.10 (s, 1H), 8.03
(s, 1H), 7.95 (s, 1H), 6.98 (s, 1H), 3.85 (s, 3H), 3.67-3.53 (m,
8H).
Example 42
5-((6-Amino-5-cyanopyrimidin-4-yl) amino)-6-methoxy-1H-indazole
##STR00075##
[0416] Step 1
5-((6-Amino-5-bromopyrimidin-4-yl) amino)-6-methoxy-1H-indazole
[0417] N-(5-bromo-6-chloropyrimidin-4-yl)acetamide 42a (84.0 mg,
0.4 mmol), 1-t-butoxycarbonyl-5-amino-6-methoxyindazole (105.0 mg,
0.4 mmol), cesium carbonate (391.0 mg, 1.2 mmol) and
N,N-dimethylacetamide (5 mL) were mixed and stirred at 120.degree.
C. for 24 hours. Then it was cooled to room temperature and
extracted with dichloromethane (30 mL.times.3). The combined
organic layer was washed with saturated brine (30 mL), dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep HPLC (water (with 0.2% formic acid), 10%.about.50%
acetonitrile, 15 minutes) gave 5-((6-amino-5-bromopyrimidin-4-yl)
amino)-6-methoxy-1H-indazole 42b (18.0 mg, 0.05 mmol, white solid),
yield: 25%.
[0418] MS m/z (ESI): 335 & 337 [M+1].
[0419] Step 2
5-((6-Amino-5-cyanopyrimidin-4-yl) amino)-6-methoxy-1H-indazole
[0420] To the mixture of 5-((6-amino-5-bromopyrimidin-4-yl)
amino)-6-methoxy-1H-indazole 42b (10.0 mg, 0.4 mmol), zinc cyanide
(7.0 mg, 0.06 mmol) and N,N-dimethylacetamide (1 mL) were added
copper(I) iodide (5.0 mg, 0.03 mmol) and
tetrakis(triphenylphosphine)palladium(0) under argon. The mixture
was reacted at 150.degree. C. for 1 hour under argon in microwave
oven. Then it was cooled to room temperature and filtered. The
filtrate was concentrated. Purification of the residue using prep
HPLC (water (0.2% formic acid), 20%.about.60% acetonitrile, 15
minutes) gave 5-((6-amino-5-cyanopyrimidin-4-yl)
amino)-6-methoxy-1H-indazole 42 (3.0 mg, 0.01 mmol, dark yellow
solid). Yield: 36%.
[0421] MS m/z (ESI): 282 [M+1].
[0422] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.52 (s, 1H), 8.25
(s, 1H), 8.07 (s, 1H), 6.94 (s, 1H), 4.98 (s, 2H), 3.90 (s,
3H).
Example 43&44
1-tert-Butoxycarbonyl-5-((6-amino-5-(pyridin-2-yl) pyrimidin-4-yl)
amino-6-methoxyindazole 5-((6-amino-5-(pyridin-2-yl)
pyrimidin-4-yl) amino-6-methoxy-1H-indazole formate
##STR00076## ##STR00077##
[0424] Step 1
4,6-Diamino-5-(4,4,5,5-tetramethyl-1,3,2-dioxoborolan-2-yl)
pyrimidine
[0425] To the mixture of 4,6-diamino-5-iodopyrimidine 43a (125.0
mg, 0.5 mmol), bis(pinacolato)diboron (127.0 mg, 0.5 mmol) and
1,4-dioxane (5 mL) was added cesium carbonate (500.0 mg, 1.5 mmol),
palladium(II) acetate (11.0 mg, 0.05 mmol) and
2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (41.0 mg, 0.1 mmol)
at room temperature under argon. The mixture was stirred at
60.degree. C. for 16 hours under argon. Then it was quenched with
water (10 mL). The organic layer was separated and the aqueous
layer was extracted with dichloromethane (15 mL.times.2). The
combined organic layer was washed with saturated brine (50
mL.times.2), dried over anhydrous sodium sulfate and filtered. The
filtrate was concentrated under reduced pressure to give
4,6-diamino-5-(4,4,5,5-tetramethyl-1,3,2-dioxoborolan-2-yl)
pyrimidine 43b as a crude product. It was used directly in next
step without further purification.
[0426] MS m/z (ESI): 237 [M+1].
[0427] Step 2
4,6-Diamino-5-(pyridin-2-yl)pyrimidine
[0428] To the mixture of
4,6-diamino-5-(4,4,5,5-tetramethyl-1,3,2-dioxoborolan-2-yl)
pyrimidine 43b (118.0 mg, 0.5 mmol), 2-bromopyridine (30.0 mg, 0.2
mmol) and 1,4-dioxane (5 mL) were added potassium acetate (98.0 mg,
1 mmol), palladium(II) acetate (9.0 mg, 0.01 mmol) and
2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (8.0 mg, 0.02 mmol)
at room temperature under argon. The mixture was stirred at
100.degree. C. for 16 hours under argon. Then it was quenched with
water (10 mL). The organic layer was separated and the aqueous
layer was extracted with dichloromethane (15 mL.times.2). The
combined organic layer was washed with saturated brine (50
mL.times.2), dried over anhydrous sodium sulfate and filtered. The
filtrate was concentrated under reduced pressure. Purification of
the residue using Prep HPLC (water (0.2% formic acid), 0%.about.10%
acetonitrile, 15 minutes) gave
4,6-diamino-5-(pyridin-2-yl)pyrimidine 43c (10.0 mg, 0.05 mmol,
yellow solid). Yield: 10%.
[0429] MS m/z (ESI): 188 [M+1].
[0430] Step 3
1-tert-Butoxycarbonyl-5-((6-amino-5-(pyridin-2-yl) pyrimidin-4-yl)
amino-6-methoxyindazole & 5-((6-Amino-5-(pyridin-2-yl)
pyrimidin-4-yl) amino-6-methoxy-1H-indazole
[0431] To the mixture of 4,6-diamino-5-(pyridin-2-yl)pyrimidine 43c
(10.0 mg, 0.05 mmol), 1-tert-butoxyacyl-5-bromo-6-methoxyindazole
(16.0 mg, 0.05 mmol) and 1,4-dioxane (5 mL) were added cesium
carbonate (32 mg, 0.1 mmol),
tris(dibenzylideneacetone)dipalladium(0) (5 mg, 0.005 mmol) and
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (6 mg, 0.01 mmol)
at room temperature under argon. The mixture was stirred at
120.degree. C. for 16 hours under argon. Then it was quenched with
water (10 mL). The organic layer was separated and the aqueous
layer was extracted with dichloromethane (15 mL.times.2). The
combined organic layer was washed with saturated brine (50
mL.times.2), dried over anhydrous sodium sulfate and filtered. The
filtrate was concentrated under reduced pressure. Purification of
the residue using Prep HPLC (water (with 0.8% ammonia bicarbonate),
20%-70% acetonitrile, 15 minutes)
gavel-tert-butoxycarbonyl-5-((6-amino-5-(pyridin-2-yl)
pyrimidin-4-yl) amino-6-methoxyindazole 43 (1.0 mg, 0.02 mmol,
white solid). Yield: 4%. Further purification of the residue using
Prep HPLC (water (with 0.2% formic acid), 20%.about.70%
acetonitrile, 15 minutes) gave 5-((6-amino-5-(pyridin-2-yl)
pyrimidin-4-yl) amino-6-methoxy-1H-indazole formate 44 (1.0 mg,
0.003 mmol, white solid). Yield: 6%.
[0432] MS m/z (ESI): 434 [M+1],
[0433] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.58 (s, 1H),
8.98 (s, 1H), 8.56 (d, J=4.4 Hz, 1H), 8.31 (s, 1H), 8.22 (s, 2H),
7.95-7.93 (m, 1H), 7.82 (d, J=8.4 Hz, 1H), 7.58 (s, 1H), 7.43-7.39
(m, 1H), 6.59 (s, 1H), 4.00 (s, 3H), 2.09 (s, 9H).
[0434] MS m/z (ESI): 334 [M+1],
[0435] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.78 (s, 1H), 10.29
(s, 1H), 8.81 (s, 1H), 8.28 (s, 1H), 8.17 (s, 2H), 7.94-7.91 (m,
2H), 7.80-7.78 (d, J=7.2 Hz, 1H), 7.40-7.38 (d, J=5.6 Hz, 1H), 6.96
(s, 1H), 6.49 (s, 2H), 3.91 (s, 3H).
Example 45
5-((6-Amino-5-fluoropyrimidin-4-yl)amino)-6-methoxy-1H-indazole
##STR00078##
[0437] Step 1
6-Chloro-5-fluoropyrimidin-4-amine
[0438] 4,6-Dichloro-5-fluoropyrimidine 45a (0.5 g, 3.0 mmol),
aqueous ammonia (3 mL) and butan-1-ol (2 mL) were mixed and stirred
at 90.degree. C. for 3 hours. White solid was formed and filtered.
The filter cake was washed with acetonitrile (50 mL) and dried to
give 6-chloro-5-fluoropyrimidin-4-amine 45b (0.2 g, 0.14 mmol,
white solid). Yield: 48%.
[0439] MS m/z (ESI): 148 & 150 [M+1].
[0440] Step 2
1-((2-(Trimethylsilyl) ethoxy)
methyl)-5-((6-amino-5-fluoropyrimidin-4-yl)
amino)-6-methoxyindazole
[0441] To the mixture of 6-chloro-5-fluoropyrimidin-4-amine 45b
(30.0 mg, 0.2 mmol),
6-methoxy-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-indazol-5-amine
(32.0 mg, 0.1 mmol) and N,N-dimethylacetamide (2 mL) were added
tris(dibenzylidene-acetone)dipalladium(0) (9.0 mg, 0.01 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (12 mg, 0.02 mmol)
and cesium carbonate (98 mg, 0.3 mmol) at room temperature under
argon. The mixture was stirred at 125.degree. C. for 1 hour under
argon in microwave oven. Then it was cooled to room temperature and
extracted with dichloromethane (20 mL.times.3). The combined
organic layer was washed with brine (20 mL), dried over anhydrous
sodium sulfate and filtered. The filtrate was concentrated under
reduced pressure. Purification of the residue using Prep HPLC
(water (with 0.2% formic acid), 10%.about.50% acetonitrile, 15
minutes) gave 1-((2-(trimethylsilyl) ethoxy)
methyl)-5-((6-amino-5-fluoropyrimidin-4-yl)
amino)-6-methoxyindazole 45c (6.0 mg, 0.015 mmol, white solid).
Yield: 15%.
[0442] MS m/z (ESI): 405 [M+1].
[0443] Step 3
5-((6-Amino-5-fluoropyrimidin-4-yl)
amino)-6-methoxy-1H-indazole
[0444] 1-((2-(Trimethylsilyl) ethoxy)
methyl)-5-((6-amino-5-fluoropyrimidin-4-yl)
amino)-6-methoxyindazole 45c (6.0 mg, 0.015 mmol), dichloromethane
(2 mL) and trifluoroacetic acid (1 mL) were mixed and stirred at
room temperature for 1 hour. Then it was quenched by saturated
aqueous sodium bicarbonate (5 mL) and diluted with dichloromethane
(5 mL). The organic layer was separated and the aqueous layer was
extracted with dichloromethane (10 mL.times.3). The combined
organic layer was washed with brine (10 mL), dried over anhydrous
sodium sulfate and filtered. The filtrate was concentrated under
reduced pressure. Purification of the residue using Prep HPLC
(water (with 0.2% formic acid), 30%.about.70% acetonitrile, 15
minutes) gave 5-((6-amino-5-fluoropyrimidin-4-yl)
amino)-6-methoxy-1H-indazole 45 (2.0 mg, 0.007 mmol, white solid).
Yield: 50%.
[0445] MS m/z (ESI): 275 [M+1],
[0446] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.83 (s, 1H), 8.15
(s, 1H), 7.94 (s, 1H), 7.80 (s, 1H), 7.77 (s, 1H), 7.02 (s, 1H),
6.63 (s, 2H), 3.89 (s, 3H).
Example 46
5-((6-Amino-5-methoxypyrimidin-4-yl)
amino)-6-methoxy-1H-indazole
##STR00079##
[0448] The synthetic procedure is similar to that of example 45.
Using 6-chloro-5-methoxypyrimidine-4-amine instead of
5-fluorinepyrimidin-4-amine. The desired compound:
5-((6-amino-5-methoxypyrimidin-4-yl) amino)-6-methoxy-1H-indazole
46 (2.0 mg, 0.007 mmol, white solid) was obtained. Yield: 27%.
[0449] MS m/z (ESI): 287 [M+1],
[0450] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.87 (s, 1H), 8.56
(s, 1H), 7.94 (s, 1H), 7.89 (s, 1H), 7.60 (s, 1H), 7.04 (s, 1H),
6.39 (s, 2H), 3.96 (s, 3H), 3.69 (s, 3H).
Example 47
5-((6-Amino-5-methylpyrimidin-4-yl)
amino)-6-methoxy-1H-indazole
##STR00080##
[0452] The synthetic procedure is similar to that of example 45.
Using 6-chloro-5-methylpyrimidine-4-amine instead of
5-fluorinepyrimidine-4-amine. The desired compound:
5-((6-amino-5-methylpyrimidin-4-yl) amino)-6-methoxy-1H-indazole 47
(1.0 mg, 0.004 mmol, white solid) was obtained. Yield: 36%.
[0453] MS m/z (ESI): 271 [M+1],
[0454] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.75 (s, 1H), 8.32
(s, 1H), 8.00-7.81 (m, 2H), 7.27 (s, 1H), 7.00 (s, 1H), 6.15 (s,
2H), 3.91 (s, 3H), 1.96 (s, 3H).
Example 48
5-((6-(Cyclopropionamido)-5-methylpyrimidin-4-yl)
amino)-6-methoxy-1H-indazole
##STR00081##
[0456] Step 1
[0457] N-(6-Chloro-5-methylpyrimidin-4-yl)cyclopropanecarboxamide
6-Chloro-5-methylpyrimidin-4-amine 48a (100.0 mg, 0.7 mmol),
cyclopropylformyl chloride (87 mg, 0.839 mmol), pyridine (3 mL) and
tetrahydrofuran (3 mL) were mixed at room temperature and stirred
at 60.degree. C. for 16 hours. Then it was diluted with water (20
mL) and extracted with ethyl acetate (20 mL.times.2). The organic
layer was washed with brine (20 mL.times.2), dried over anhydrous
sodium sulfate and filtered. The filtrate was concentrated under
reduced pressure. Purification of the residue using Prep TLC
(petroleum ether/ethyl acetate=4:1) gave
N-(6-chloro-5-methylpyrimidin-4-yl)cyclopropanecarboxamide 48b
(40.0 mg, 0.19 mmol, white solid), yield: 27%.
[0458] MS m/z (ESI): 212 & 214 [M+1];
[0459] Step 2
1-((2-(Trimethylsilyl) ethoxy)
methyl)-5-((6-(cyclopropionylamino)-5-methylpyrimidin-4-yl)
amino)-6-Methoxyindazole
[0460] To the solution of
N-(6-chloro-5-methylpyrimidin-4-yl)cyclopropanecarboxamide 48b
(10.0 mg, 0.05 mmol),
6-methoxy-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-indazol-5-amine
(20.0 mg, 0.071 mmol) and cesium carbonate (46 mg, 0.142 mmol) in
1,4-dioxane (1 mL) were added
tris(dibenzylideneacetone)dipalladium(0) (4 mg, 0.005 mmol) and
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (5 mg, 0.009 mmol)
at room temperature under argon. The mixture was stirred at
110.degree. C. for 1 hour under argon in microwave oven. Then it
was diluted with water (10 mL) and extracted with ethyl acetate (10
mL.times.3). The combined organic layer was washed with brine (10
mL.times.2), dried over anhydrous sodium sulfate and filtered. The
filtrate was concentrated under reduced pressure. Purification of
the residue using Prep TLC (petroleum ether/ethyl acetate=3:2) gave
1-((2-(trimethylsilyl) ethoxy)
methyl)-5-((6-(cyclopropionylamino)-5-methylpyrimidin-4-yl)
amino)-6-Methoxyindazole 48c (10.0 mg, 0.021 mmol, pale yellow
solid).
[0461] Yield: 46%.
[0462] MS m/z (ESI): 469 [M+1].
[0463] Step 3
5-((6-(Cyclopropionamido)-5-methylpyrimidin-4-yl)
amino)-6-methoxy-1H-indazole
[0464] 1-((2-(Trimethylsilyl) ethoxy)
methyl)-5-((6-(cyclopropionylamino)-5-methylpyrimidin-4-yl)
amino)-6-Methoxyindazole 48c (10.0 mg, 0.021 mmol), dichloromethane
(0.5 mL) and trifluoroacetic acid (1 mL) were mixed and stirred at
room temperature for 1 hour. Then it was diluted with water (10 mL)
and extracted with dichloromethane (10 mL.times.3). The combined
organic layer was dried over anhydrous sodium sulfate and filtered.
The filtrate was concentrated under reduced pressure. Purification
of the residue using Prep HPLC (water (with 0.8% ammonia
bicarbonate), 40%.about.60% acetonitrile, 15 minutes) gave
5-((6-(cyclopropionamido)-5-methylpyrimidin-4-yl)
amino)-6-methoxy-1H-indazole 48 (3.0 mg, 0.009 mmol, white solid).
Yield: 42%.
[0465] MS m/z (ESI): 339 [M+1],
[0466] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.85 (s, 1H), 10.29
(s, 1H), 8.21 (s, 1H), 8.04 (s, 1H), 7.99 (s, 1H), 7.96 (s, 1H),
7.04 (s, 1H), 3.86 (s, 3H), 1.96 (s, 3H), 1.95-1.92 (m, 1H),
0.83-0.81 (m, 4H).
Example 49
5-((6-(Cyclopropanesulfonylamino) pyrimidin-4-yl)
amino)-6-methoxy-1H-indazole
##STR00082##
[0468] Step 1
1-tert-Butoxyacyl-5-((6-(cyclopropanesulfonylamino) pyrimidin-4-yl)
amino)-6-methoxyindazole
[0469] To the mixture of
N-(6-aminopyrimidin-4-yl)cyclopropanesulfonamide 49a (21.0 mg, 0.1
mmol), 1-tert-butoxyacyl-5-bromo-6-methoxyindazole (33.0 mg, 0.1
mmol) and 1,4-dioxane (2 mL) were added
tris(dibenzylideneacetone)-dipalladium(0) (9.0 mg, 0.01 mmol),
cesium carbonate (65.0 mg, 0.2 mmol) and
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (12.0 mg, 0.02
mmol) at room temperature under argon. The mixture was stirred at
110.degree. C. for 15 hours under argon. Then it was quenched with
water (10 mL). The organic layer was separated and the aqueous
layer was extracted with dichloromethane (15 mL.times.2). The
combined organic layer was washed with brine (50 mL.times.2), dried
over anhydrous sodium sulfate and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep TLC (dichloromethane/methanol=20:1) gave
1-tert-butoxyacyl-5-((6-(cyclopropanesulfonylamino) pyrimidin-4-yl)
amino)-6-methoxyindazole 49b (20.0 mg, 0.043 mmol, yellow solid).
Yield: 43%.
[0470] MS m/z (ESI): 461 [M+1].
[0471] Step 2
5-((6-(Cyclopropanesulfonylamino) pyrimidin-4-yl)
amino)-6-methoxy-1H-indazole
[0472] 1-tert-Butoxyacyl-5-((6-(cyclopropanesulfonylamino)
pyrimidin-4-yl) amino)-6-methoxyindazole 49b (20.0 mg, 0.043 mmol),
dichloromethane (1.0 mL) and trifluoroacetic acid (1.0 mL) were
mixed and stirred at room temperature for 3 hours. Then it was
quenched by saturated aqueous sodium bicarbonate solution (10 mL).
The organic layer was separated and the aqueous layer was extracted
with dichloromethane (15 mL.times.2). The combined organic layer
was washed with brine (50 mL.times.2), dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated under reduced
pressure. Purification of the residue using Prep TLC
(dichloromethane/methaol=10:1) gave
5-((6-(cyclopropanesulfonylamino) pyrimidin-4-yl)
amino)-6-methoxy-1H-indazole 49 (2.0 mg, 0.006 mmol, white solid).
Yield: 14%.
[0473] MS m/z (ESI): 361 [M+1];
[0474] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.89 (s, 1H), 8.98
(s, 1H), 8.32 (s, 1H), 8.16 (s, 1H), 7.95 (s, 1H), 7.80 (s, 1H),
7.04 (s, 1H), 6.15 (s, 1H), 3.78 (s, 3H), 2.67-2.56 (m, 1H),
0.90-0.88 (m, 4H).
Example 50
5-(6-(Acetamido) pyrimidin-4-yl) amino)-6-methoxy-1H-pyrazolo
[3,4-b] pyridine formate
##STR00083##
[0476] Step 1
6-Chloro-1H-pyrazolo[3,4-b]pyridine
[0477] To the solution of 2,6-dichloronicotinaldehyde 50a (4.0 g,
22.86 mmol) in butan-1-ol (1 mL) was added hydrazine hydrate (3.3
mL, 68.57 mmol) at room temperature. The mixture was stirred at
120.degree. C. for 8 hours. Then it was concentrated in vacuum. The
residue was diluted with water (100 mL) and extracted with ethyl
acetate (100 mL.times.3). The combined organic layer was washed
with brine (100 mL.times.2), dried over anhydrous sodium sulfate
and filtered.
[0478] The filtrate was concentrated under reduced pressure.
Purification of the residue using flash column chromatography
(petroleum ether.about.petroleum ether/ethyl acetate=7:1) gave
6-chloro-1H-pyrazolo[3,4-b]pyridine 50b (0.75 g, 4.90 mmol, yellow
solid). Yield: 21%.
[0479] MS m/z (ESI): 154 & 156 [M+1];
[0480] Step 2
1-Benzyl-6-chloro-1H-pyrazolo[3,4-b]pyridine
[0481] 6-Chloro-1H-pyrazolo[3,4-b]pyridine 50b (0.60 g, 3.92 mmol),
benzyl bromide (738 mg, 4.31 mmol), cesium carbonate (1.4 g, 4.31
mmol) and N,N-dimethylformamide (3 mL) were mixed and stirred at
room temperature for 2 hours. Then it was quenched with saturated
aqueous sodium thiosulfate solution (50 mL) and extracted with
ethyl acetate (50 mL.times.2). The combined organic layer was
washed with brine (50 mL.times.2), dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated under reduced
pressure. Purification of the residue using flash column
chromatography (petroleum ether-petroleum ether/ethyl
acetate=100:0-100:2) gave
1-benzyl-6-chloro-1H-pyrazolo[3,4-b]pyridine 50c (0.75 g, 3.09
mmol, yellow solid), yield: 27%.
[0482] Step 3
1-Benzyl-1,7-dihydro-6H-pyrazolo[3,4-b]pyridin-6-one
[0483] To the solution of
1-benzyl-6-chloro-1H-pyrazolo[3,4-b]pyridine 50c (0.75 g, 3.09
mmol) in water (8 mL) and dimethyl sulfoxide (8 mL) was added
sodium hydroxide (1.2 g, 30.86 mmol). The resulting mixture was
stirred at 100.degree. C. for 16 hours. Then it was cooled to room
temperature and adjusted to pH a 8 with concentrated hydrochloric
acid. The mixture was diluted with water (50 mL) and extracted with
ethyl acetate (50 mL.times.3). The combined organic layer was
washed with water (50 mL.times.3) and saturated brine (50
mL.times.2), dried over anhydrous sodium sulfate and filtered. The
filtrate was concentrated under reduced pressure to give
1-benzyl-1,7-dihydro-6H-pyrazolo[3,4-b]pyridin-6-one 50d (0.6 g,
2.67 mmol, yellow solid). Yield: 86%.
[0484] MS m/z (ESI): 226 [M+1],
[0485] Step 4
1-Benzyl-5-bromo-1,7-dihydro-6H-pyrazolo[3,4-b]pyridin-6-one
[0486] To the solution of
1-benzyl-1,7-dihydro-6H-pyrazolo[3,4-b]pyridin-6-one 50d (0.38 g,
1.69 mmol) in water (6 mL) and acetonitrile (8 mL) was added
lithium hydroxide (81 mg, 3.38 mol) and N-bromosuccinimide (0.60 g,
3.38 mol). The mixture was stirred at room temperature for 2 hours.
Then it was quenched with saturated aqueous sodium thiosulfate
solution (20 mL) and extracted with ethyl acetate (20 mL.times.3).
The combined organic layer was washed with brine (20 mL.times.2),
dried over anhydrous sodium sulfate and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using flash column chromatography (petroleum ether/ethyl
acetate=3:1.about.2:3) gave
1-benzyl-5-bromo-1,7-dihydro-6H-pyrazolo[3,4-b]pyridin-6-one 50e
(0.23 g, 0.76 mmol, pale yellow solid). Yield: 45%.
[0487] MS m/z (ESI): 304 & 306 [M+1],
[0488] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.16 (s, 1H), 7.76
(s, 1H), 7.52-7.29 (m, 5H), 5.61 (s, 2H).
[0489] Step 5
1-Benzyl-5-bromo-6-methoxy-1H-pyrazolo[3,4-b]pyridine
[0490] 1-Benzyl-5-bromo-1,7-dihydro-6H-pyrazolo[3,4-b]pyridin-6-one
50e (0.23 g, 0.76 mmol), potassium carbonate (0.21 g, 1.52 mmol),
iodomethane (0.22 g, 1.518 mmol) and N,N-dimethylformamide (4 mL)
were mixed and stirred at room temperature for 2 hours. Then it was
diluted with water (20 mL) and extracted with ethyl acetate (20
mL.times.3). The combined organic layer was washed with water (20
mL.times.3) and saturated brine (20 mL.times.2), dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using flash column chromatography (petroleum ether/ethyl
acetate=1:0-25:1) gave
1-benzyl-5-bromo-6-methoxy-1H-pyrazolo[3,4-b]pyridine 50f (95 mg,
0.30 mmol, pale yellow solid). Yield: 39%.
[0491] MS m/z (ESI): 318 & 320 [M+1],
[0492] Step 6
N-(6-((1-Benzyl-6-methoxy-1H-pyrazolo[3,4-b]pyridin-5-yl)amino)pyrimidin-4-
-yl)acetamide
[0493] To the solution of
1-benzyl-5-bromo-6-methoxy-1H-pyrazolo[3,4-b]pyridine 50f (20.0 mg,
0.063 mmol), N-(6-aminopyrimidin-4-yl)acetamide (19.0 mg, 0.126
mmol) and cesium carbonate (62 mg, 0.189 mmol) in 1,4-dioxane (1
mL) was added tris(dibenzylideneacetone)dipalladium(0) (6 mg, 0.006
mmol) and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (7 mg,
0.013 mmol) at room temperature under argon. The mixture was
stirred at 100.degree. C. for 1 hour under argon in microwave oven.
Then it was diluted with water (10 mL) and extracted with ethyl
acetate (10 mL.times.3). The combined organic layer was washed with
brine (10 mL.times.2), dried over anhydrous sodium sulfate and
filtered. The filtrate was concentrated under reduced pressure.
Purification of the residue using Prep TLC (petroleum ether/ethyl
acetate=3:1) gave
N-(6-((1-benzyl-6-methoxy-1H-pyrazolo[3,4-b]pyridin-5-yl)amino)pyrimidin--
4-yl)acetamide 50g (10.0 mg, 0.026 mmol, pale yellow solid). Yield:
41%.
[0494] MS m/z (ESI): 390 [M+1].
[0495] Step 7
5-(6-(Acetamido) pyrimidin-4-yl) amino)-6-methoxy-1H-pyrazolo
[3,4-b] pyridine formate
[0496] To the solution of
N-(6-((1-benzyl-6-methoxy-1H-pyrazolo[3,4-b]pyridin-5-yl)amino)pyrimidin--
4-yl)acetamide 50g (10.0 mg, 0.026 mmol) in tetrahydrofuran (2 mL)
was added dropwise methyllithium (0.16 mL, 0.257 mmol, 2 M) under
argon. The mixture was stirred at room temperature for 2 hours.
Then it was quenched with water (10 mL) and extracted with
dichloromethane (10 mL.times.3). The combined organic layer was
dried over anhydrous sodium sulfate and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep HPLC (water (with 0.2% formic acid), 10%.about.60%
acetonitrile, 15 minutes) gave 5-(6-(acetamido) pyrimidin-4-yl)
amino)-6-methoxy-1H-pyrazolo [3,4-b] pyridine formate 50 (1.1 mg,
0.003 mmol, white solid). Yield: 13%.
[0497] MS m/z (ESI): 300 [M+1],
[0498] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 13.27 (s, 1H), 10.38
(s, 1H), 8.91 (s, 1H), 8.43 (s, 1H), 8.40 (s, 1H), 8.27 (s, 1H),
7.98 (s, 1H), 7.46 (s, 1H), 3.87 (s, 3H), 2.07 (s, 3H).
Example 51
5-(6-Methoxypyrimidin-4-yl) amino)-6-methoxy-1H-indazole
formate
##STR00084##
[0500] Step 1
1-((2-(Trimethylsilyl) ethoxy) methyl)-5-(6-methoxypyrimidin-4-yl)
amino)-6-methoxyindazole
[0501] To the mixture of 1-((2-(trimethylsilyl) ethoxy)
methyl)-5-amino-6-methoxyindazole 51a (30.0 mg, 0.10 mmol),
4-chloro-6-methoxypyrimidine (14.4 mg, 0.10 mmol) and 1, 4-dixane
(1 mL) were added tris(dibenzylideneacetone)dipalladium(0) (9.2 mg,
0.01 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (5.8
mg, 0.01 mmol) and cesium carbonate (65.2 mg, 0.2 mmol) at room
temperature under argon. The mixture was stirred at 120.degree. C.
for 1 hour under argon in microwave oven. Then it was cooled to
room temperature, diluted with dichloromethane (10 mL) and
filtered. The filtrate was concentrated under reduced pressure.
Purification of the residue using Prep HPLC (water (with 0.2%
formic acid), 60%.about.80% acetonitrile, 15 minutes) gave
1-((2-(trimethylsilyl) ethoxy) methyl)-5-(6-methoxypyrimidin-4-yl)
amino)-6-methoxyindazole 51b (4.6 mg, 0.011 mmol, white solid).
Yield: 12%.
[0502] MS m/z (ESI): 402 [M+1].
[0503] Step 2
5-(6-Methoxypyrimidin-4-yl) amino)-6-methoxy-1H-indazole
formate
[0504] 1-((2-(Trimethylsilyl) ethoxy)
methyl)-5-(6-methoxypyrimidin-4-yl) amino)-6-methoxyindazole 51b
(4.6 mg, 0.011 mmol) was dissolved into dichloromethane (1 mL) and
trifluoroacetic acid (1 mL) was added, and the resultant mixture is
stirred at room temperature for 2 hours. Then it was concentrated
under reduced pressure. The residue was dissolved in dimethyl
sulfoxide (1 mL). The mixture was adjusted to pH=8.about.9 with
saturated aqueous sodium hydroxide solution. Purification of the
mixture using Prep HPLC (water (with 0.2% formic acid),
10%.about.40% acetonitrile, 15 minutes) gave
5-(6-methoxypyrimidin-4-yl) amino)-6-methoxy-1H-indazole formate 51
(1.5 mg, 0.006 mmol, white solid). Yield: 55%.
[0505] MS m/z (ESI): 272 [M+1],
[0506] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.89 (s, 1H), 8.61
(s, 1H), 8.28 (s, 1H), 8.24 (s, 1H), 8.02 (s, 1H), 7.94 (s, 1H),
7.03 (s, 1H), 5.97 (s, 1H), 3.87 (s, 3H), 3.80 (s, 3H).
Example 52
5-(5-Methoxypyrimidin-4-yl) amino)-6-methoxy-1H-indazole
##STR00085##
[0508] The synthetic procedures were similar to those in Example
49. Using 5-methoxypyrimidine-4-amine instead of
N-(6-aminopyrimidine-4-yl)cyclopropylsulfonamide, the desired
compound 5-(5-methoxypyrimidin-4-yl) amino)-6-methoxy-1H-indazole
52 (9.0 mg, 0.003 mmol, white solid) was obtained. Yield: 47%.
[0509] MS m/z (ESI): 272 [M+1],
[0510] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.84 (s, 1H), 8.71
(s, 1H), 8.33 (s, 1H), 8.08 (s, 1H), 8.05 (s, 1H), 7.99 (s, 1H),
7.08 (s, 1H), 3.98 (s, 3H), 3.96 (s, 3H).
Example 53
5-(5-(2-Methoxyethoxy) pyrimidin-4-yl)
amino)-6-methoxy-1H-indazole
##STR00086##
[0512] Step 1
4-Amino-5-hydroxypyrimidine
[0513] To the solution of 4-amino-5-methoxypyrimidine 53a (500.0
mg, 4.0 mmol) and sodium methanolate (432.0 mg, 8.0 mmol) in
anhydrous N,N-dimethylformamide (6 mL) was added 1-dodecanethiol
(1.61 g, 8.00 mmol). The mixture was stirred at 120.degree. C. for
12 hours. Then it was cooled to room temperature and concentrated
under reduced pressure. The residue was adjusted pH to 5.about.6
with 5.0 mL of water and 0.5 mL of acetic acid. The aqueous layer
was washed with ethyl acetate (10 mL.times.5) and concentrated in
vacuum to give 4-amino-5-hydroxypyrimidine 53b (0.4 g, 3.6 mmol,
off-white solid), yield: 90%.
[0514] MS m/z (ESI): 112 [M+1],
[0515] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.64 (brs, 1H), 7.91
(s, 1H), 7.63 (s, 1H), 6.42 (brs, 2H).
[0516] Step 2
5-(2-Methoxyethoxy)pyrimidin-4-amine
[0517] The solution of 4-amino-5-hydroxypyrimidine 53b (100.0 mg,
0.9 mmol) and anhydrous lithium hydroxide (65.0 mg, 2.7 mmol) in
N,N-dimethylformamide (6 mL) was stirred at room temperature for 30
minutes. 1-Bromo-2-methoxyethane (125.0 mg, 0.9 mmol) was added.
The reaction mixture was stirred at 60.degree. C. for 5 hours. Then
it was cooled to room temperature and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep TLC (dichloromethane/methanol=10:1) gave
5-(2-methoxyethoxy)pyrimidin-4-amine 53c (60.0 mg, 0.36 mmol, clear
oil), yield: 40%.
[0518] MS m/z (ESI): 170 [M+1],
[0519] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.16 (s, 1H), 7.79
(s, 1H), 5.52 (brs, 2H), 4.10-4.12 (m, 2H), 3.67-3.69 (m, 2H), 3.37
(s, 3H).
[0520] Step 3
1-tert-Butoxycarbonyl-5-(5-(2-methoxyethoxy) pyrimidin-4-yl)
amino)-6-methoxyindazole
[0521] To the mixture of 5-(2-methoxyethoxy)pyrimidin-4-amine 53c
(30.0 mg, 0.17 mmol),
1-tert-butoxycarbonyl-5-amino-6-methoxyindazole (110.0 mg, 0.34
mmol) and cesium carbonate (180 mg, 0.51 mmol) in 1,4-dioxane (2
mL) were added tris(dibenzylideneacetone)dipalladium(0) (15.0 mg,
0.016 mmol) and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (18
mg, 0.032 mmol) at room temperature under argon. The mixture was
stirred at 120.degree. C. for 1 hour under argon in microwave oven.
Then it was cooled to room temperature and filtered. The filtrate
was concentrated under reduced pressure. Purification of the
residue using Prep TLC (dichloromethane/methanol=10:1) gave
1-tert-butoxycarbonyl-5-(5-(2-methoxyethoxy) pyrimidin-4-yl)
amino)-6-methoxyindazole 53d (13.0 mg, 0.031 mmol, white solid).
Yield: 18%.
[0522] MS m/z (ESI): 416 [M+1],
[0523] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.08 (s, 1H), 8.49
(s, 1H), 8.18 (s, 1H), 8.13 (s, 1H), 7.98 (s, 1H), 7.76 (s, 1H),
4.08-4.05 (m, 2H), 3.96 (s, 3H), 3.86-3.83 (m, 2H), 3.49 (s, 3H),
1.73 (s, 9H).
[0524] Step 4
5-(5-(2-Methoxyethoxy) pyrimidin-4-yl)
amino)-6-methoxy-1H-indazole
[0525] To the solution of
1-tert-butoxycarbonyl-5-(5-(2-methoxyethoxy) pyrimidin-4-yl)
amino)-6-methoxyindazole 53d (13.0 mg, 0.031 mmol) in
dichloromethane (1 mL) was added the solution of hydrogen chloride
in 1,4-dioxane (4 M, 1.0 mL). The mixture was stirred at room
temperature for 12 hours. Then it was concentrated under reduced
pressure. Purification of the residue using Prep HPLC (water (with
0.2% formic acid), 20%.about.40% acetonitrile, 15 minutes) gave
5-(5-(2-methoxyethoxy) pyrimidin-4-yl) amino)-6-methoxy-1H-indazole
53 (6.0 mg, 0.019 mmol, white solid). Yield: 61%.
[0526] MS m/z (ESI): 316 [M+1],
[0527] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.87 (brs, 1H), 8.77
(s, 1H), 8.36 (s, 1H), 8.15 (s, 1H), 8.11 (s, 1H), 7.99 (s, 1H),
7.09 (s, 1H), 4.33-4.30 (m, 2H), 3.96 (s, 3H), 3.78-3.75 (m, 2H),
3.40 (s, 3H).
Example 54
5-(5-(2-Hydroxyethoxy) pyrimidin-4-yl) amino)-6-methoxy-1H-indazole
formate
##STR00087##
[0529] The synthetic procedures were similar to that in Example 53.
Using (2-bromoethoxy)-t-butyldimethylsilane instead of
1-bromine-2-methoxyethane. The desired compound:
5-(5-(2-hydroxyethoxy) pyrimidin-4-yl) amino)-6-methoxy-1H-indazole
formate 54 (1.0 mg, 0.003 mmol, white solid) was obtained. Yield:
28%. Purification condition: prep HPLC (water (with 0.2% formic
acid), 10%.about.60% acetonitrile, 15 minutes).
[0530] MS m/z (ESI): 302 [M+1],
[0531] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.88 (s, 1H), 8.55
(s, 1H), 8.28-8.23 (m, 3H), 8.07 (s, 1H), 7.99 (s, 1H), 7.08 (s,
1H), 5.32 (br, 1H), 4.21-4.19 (m, 2H), 3.94 (s, 3H), 3.82-3.79 (m,
2H).
Example 55
5-(5-(2-(4-Morpholine) ethoxy) pyrimidin-4-yl)
amino)-6-methoxy-1H-indazole formate
##STR00088##
[0533] The synthetic procedures were similar to that in Example 53.
Using 4-(2-chloroethyl)morpholine instead of
1-bromine-2-methoxyethane. The desired compound:
5-(5-(2-(4-morpholine) ethoxy) pyrimidin-4-yl)
amino)-6-methoxy-1H-indazole formate 55 was obtained. Yield: 25%.
Purification condition: prep HPLC (water (with 0.2% formic acid),
10%.about.40% acetonitrile, 15 minutes).
[0534] MS m/z (ESI): 371 [M+1],
[0535] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.88 (s, 1H), 8.79
(s, 1H), 8.36 (s, 1H), 8.17 (s, 1H), 8.11 (s, 1H), 7.99 (s, 1H),
7.09 (s, 1H), 4.33-4.24 (m, 2H), 3.97 (s, 3H), 3.59-3.52 (m, 4H),
3.18-3.10 (m, 4H), 2.92-2.61 (m, 2H).
Example 56
5-(5-Ethoxypyrimidin-4-yl) amino)-6-methoxy-1H-indazole formate
##STR00089##
[0537] The synthetic procedures were similar to that in Example 53.
Using iodoethane instead of 1-bromine-2-methoxyethane. The desired
compound: 5-(5-ethoxypyrimidin-4-yl) amino)-6-methoxy-1H-indazole
formate 56 was obtained.
[0538] Yield: 25%. Purification condition: prep HPLC (water (with
0.2% formic acid), 10%.about.50% acetonitrile, 15 minutes).
[0539] MS m/z (ESI): 286 [M+1],
[0540] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.91 (brs, 1H), 8.73
(s, 1H), 8.33 (s, 1H), 8.21 (s, 1H), 8.11 (s, 1H), 8.07 (s, 1H),
7.99 (s, 1H), 7.09 (s, 1H), 4.28-4.21 (m, 2H), 3.97 (s, 3H), 1.44
(t, J=6.4, 3H).
Example 57
5-(5-Anilinopyrimidin-4-yl) amino)-6-methoxy-1H-indazole
formate
##STR00090##
[0542] Step 1
1-((2-(Trimethylsilyl) ethoxy) methyl)-5-(5-bromopyrimidin-4-yl)
amino)-6-methoxyindazole
[0543] 1-((2-(Trimethylsilyl) ethoxy)
methyl)-5-amino-6-methoxyindazole 57a (200 mg, 0.68 mmol),
5-bromo-4-chloropyrimidine (164 mg, 0.9 mmol), cesium carbonate
(445 mg, 1.4 mmol) and N,N-dimethylformamide (5 mL) were mixed and
stirred at 150.degree. C. for 1 hour in microwave oven. Then it was
diluted with dichloromethane (20 mL) and washed with brine (50
mL.times.2). The organic layer was dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated under reduced
pressure. Purification of the residue using flash column
chromatography (dichloromethane/methanol=20:1) gave
1-((2-(trimethylsilyl) ethoxy) methyl)-5-(5-bromopyrimidin-4-yl)
amino)-6-methoxyindazole 57b (102.0 mg, 0.23 mmol, yellow oil).
Yield: 34%.
[0544] MS m/z (ESI): 450 & 452 [M+1];
[0545] Step 2
1-((2-(Trimethylsilyl) ethoxy) methyl)-5-(5-anilinopyrimidin-4-yl)
amino)-6-methoxyindazole
[0546] To the mixture of 1-((2-(trimethylsilyl) ethoxy)
methyl)-5-(5-bromopyrimidin-4-yl) amino)-6-methoxyindazole 57b
(30.0 mg, 0.067 mmol), aniline (9.3 mg, 0.10 mmol) and 1,4-dioxane
(1 mL) were added tris(dibenzylideneacetone)-dipalladium(0) (6.1
mg, 0.007 mmol),
2-(dicyclohexylphosphino)3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-bipheny-
l (7.2 mg, 0.013 mmol) and cesium carbonate (87.4 mg, 0.27 mmol)
under argon. The mixture was stirred at 90.degree. C. for 1 hour
under argon in microwave oven. Then it was cooled to room
temperature, diluted with dichloromethane (10 mL) and filtered. The
filtrate was concentrated under reduced pressure. Purification of
the residue using Prep HPLC (water (with 0.2% formic acid),
30%.about.80% acetonitrile, 15 minutes) gave 1-((2-(trimethylsilyl)
ethoxy) methyl)-5-(5-anilinopyrimidin-4-yl)
amino)-6-methoxyindazole 57c (1.7 mg, 0.0033 mmol, white solid).
Yield: 5%.
[0547] MS m/z (ESI): 463 [M+1];
[0548] Step 3
5-(5-Anilinopyrimidin-4-yl) amino)-6-methoxy-1H-indazole
formate
[0549] 1-((2-(Trimethylsilyl) ethoxy)
methyl)-5-(5-anilinopyrimidin-4-yl) amino)-6-methoxyindazole 57c
(1.7 mg, 0.0033 mmol) was dissolved into hydrogen chloride in
methanol (2 M, 3.0 mL) and was stirred at 80.degree. C. for 0.5
hour. Then it was concentrated under reduced pressure. Purification
of the residue using Prep HPLC (water (with 0.2% formic acid),
20%.about.40% acetonitrile, 15 minutes) gave
5-(5-Anilinopyrimidin-4-yl) amino)-6-methoxy-1H-indazole formate 57
(1.0 mg, 0.0026 mmol, white solid). Yield: 80%.
[0550] MS m/z (ESI): 333 [M+1],
[0551] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.92 (s, 1H), 8.65
(s, 1H), 8.50 (s, 1H), 8.21 (s, 1H), 8.02 (s, 1H), 7.98 (s, 1H),
7.86 (s, 1H), 7.27-7.19 (m, 2H), 7.02 (s, 1H), 6.86-6.78 (m, 3H),
6.68 (s, 1H), 3.76 (s, 3H).
Example 58
N-(6-Methoxy-1H-indazol-5-yl)-purin-6-amine
##STR00091##
[0553] Step 1
N-(6-Methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)-9H-puri-
n-6-amine
[0554] 6-Chloro-9H-purine 58a (154.0 mg, 1.0 mmol),
6-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-amine
(293.0 mg, 1.0 mmol) and acetic acid (2 mL) were mixed and stirred
at 70.degree. C. for 1 hour. Evaporation of the solvents and the
residue was diluted with dichloromethane (20 mL). It was adjusted
to pH=8.about.10 by saturated aqueous sodium bicarbonate solution.
The mixture was extracted with dichloromethane (50 mL.times.3). The
combined organic layer was washed with brine (50 mL), dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using flash column chromatography (dichloromethane/methanol=40:1),
N-(6-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)-9H-pur-
in-6-amine 58b (50.0 mg, 0.12 mmol, white solid) was obtained.
Yield: 12%.
[0555] MS m/z (ESI): 412 [M+1];
[0556] Step 2
N-(6-Methoxy-1H-indazol-5-yl)-9H-purin-6-amine
[0557]
N-(6-Methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)--
9H-purin-6-amine 58b (50.0 mg, 0.12 mmol), dichloromethane (2 mL)
and trifluoroacetic acid (1 mL) were mixed and stirred at room
temperature for 3 hours. Then it was quenched by 5 mL of saturated
aqueous sodium bicarbonate solution and diluted with
dichloromethane (5 mL). The organic layer was separated and the
aqueous layer was extracted with dichloromethane (20 mL.times.2).
The combined organic layer was washed with brine (20 mL), dried
over anhydrous sodium sulfate and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep HPLC (water (with 0.2% formic acid), 20%.about.50%
acetonitrile, 15 minutes) gave
N-(6-methoxy-1H-indazol-5-yl)-9H-purin-6-amine 58 (30.0 mg, 0.11
mmol, white solid). Yield: 80%.
[0558] MS m/z (ESI): 282 [M+1],
[0559] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.08 (s, 1H), 8.70
(s, 1H), 8.55 (s, 1H), 8.06 (s, 1H), 7.98 (s, 1H), 7.16 (s, 1H),
3.87 (s, 3H).
Example 59
N-(6-Methoxy-1H-indazol-5-yl)-9-methyl-9H-purin-6-amine
##STR00092##
[0561] Step 1
6-Chloro-9-methyl-9H-purine
[0562] To the mixture of 6-chloro-7H-purine 59a (154.0 mg, 1.0
mmol), sodium hydride (60% in mineral oil, 120.0 mg, 5.0 mmol) and
N,N-dimethylacetamide (2 mL) was added iodomethane (426.0 mg, 3.0
mmol) at room temperature. The mixture was stirred at room
temperature for 24 hours. Then it was quenched with 5 mL of water
and extracted with dichloromethane (30 mL.times.3). The combined
organic layer was washed with brine (30 mL), dried over anhydrous
sodium sulfate and filtered. The filtrate was concentrated under
reduced pressure. Purification of the residue using flash column
chromatography gave 6-chloro-9-methyl-9H-purine 59b (80.0 mg, 0.48
mmol, white solid). Yield: 48%.
[0563] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.78 (s, 1H), 8.68
(s, 1H), 3.90 (s, 3H).
[0564] Step 2
N-(6-Methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)-9-methy-
l-9H-purin-6-amine
[0565] 6-Chloro-9-methyl-9H-purine 59b (17.0 mg, 0.1 mmol),
6-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-amine
(29.0 mg, 10.1 mmol) and acetic acid (2 mL) were mixed and stirred
at 90.degree. C. for 1 hour. Evaporation of the solvents and the
residue was diluted with dichloromethane (20 mL). The pH value was
adjusted to 8.about.10 by saturated aqueous sodium bicarbonate
solution. The organic layer was separated and the aqueous layer was
extracted with dichloromethane (10 mL.times.3). The combined
organic layer was washed with brine (10 mL), dried over anhydrous
sodium sulfate and filtered. The filtrate was concentrated under
reduced pressure. Purification of the residue using prep-TLC
(dichloromethane/methanol=20:1) gave
N-(6-methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)-9-meth-
yl-9H-purin-6-amine 59c (13.0 mg, 0.03 mmol). Yield: 30%.
[0566] MS m/z (ESI): 426 [M+1].
[0567] Step 3
N-(6-Methoxy-1H-indazol-5-yl)-9-methyl-9H-purin-6-amine
[0568]
N-(6-Methoxy-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-5-yl)--
9-methyl-9H-purin-6-amine 59c (13.0 mg, 0.03 mmol), dichloromethane
(2 mL) and trifluoroacetic acid (1 mL) were mixed and stirred at
room temperature for 3 hours. Then it was quenched by 5 mL of
saturated aqueous sodium bicarbonate solution and diluted with
dichloromethane (5 mL). The organic layer was separated and the
aqueous layer was extracted with dichloromethane (10 mL.times.3).
The combined organic layer was washed with brine (10 mL), dried
over anhydrous sodium sulfate and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep HPLC (water (with 0.2% formic acid), 20%.about.50%
acetonitrile, 15 minutes) gave
N-(6-methoxy-1H-indazol-5-yl)-9-methyl-9H-purin-6-amine 59 (6.0 mg,
0.02 mmol, white solid).
[0569] Yield: 66%.
[0570] MS m/z (ESI): 296 [M+1],
[0571] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.88 (s, 1H), 8.69
(s, 1H), 8.49 (s, 1H), 8.44 (s, 1H), 8.26 (s, 1H), 7.99 (s, 1H),
7.09 (s, 1H), 4.05 (s, 3H), 3.94 (s, 3H).
Example 60&61
1-tert-Butoxycarbonyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxy-
benzimidazole
[0572]
3-tert-Butoxycarbonyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6--
methoxybenzimidazole
##STR00093##
Step 1
4-Bromo-5-methoxy-2-nitroaniline
[0573] To the mixture of 5-methoxy-2-nitroaniline 60a (2.0 g, 11.9
mmol) with acetonitrile (30.0 mL) was added N-bromosuccinimide (2.3
g, 13.1 mmol) at room temperature. The resulting mixture was
stirred at room temperature for 3 hours. Then it was quenched with
saturated aqueous sodium sulfite (60 mL) and diluted with water
until large amount of precipitate formed. The precipitate was
filtered. The filter cake was dissolved in ethyl acetate (100 mL),
dried over anhydrous sodium sulfate and filtered. The filtrate was
concentrated to give 4-bromo-5-methoxy-2-nitroaniline 60b (2.5 g,
10.1 mmol, yellow solid) as the desired product. Yield: 85%.
[0574] MS m/z (ESI): 247 & 249 [M+1].
[0575] Step 2
4-Bromo-5-methoxybenzene-1,2-diamine
[0576] 4-Bromo-5-methoxy-2-nitroaniline 60b (0.5 g, 2.0 mmol) was
mixed with dichloromethane (10.0 mL), methanol (10.0 mL) and
saturated aqueous ammonium chloride (20.0 mL), zinc powder (1.3 g,
20.0 mmol) was added at room temperature. The resulting mixture was
stirred at room temperature for 2 hours. Then it was filtered. The
filtrate was diluted with dichloromethane (100 mL), dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated to give 4-bromo-5-methoxybenzene-1,2-diamine 60c
(450.0 mg, 2.0 mmol, brown solid). Yield: 100%.
[0577] MS m/z (ESI): 217 & 219 [M+1].
[0578] Step 3
5-Bromo-6-methoxy-1H-benzo[d]imidazole
[0579] To the mixture of 4-bromo-5-methoxybenzene-1,2-diamine 60c
(450.0 mg, 2.0 mmol) and triethoxymethane (10.0 mL) was added
formic acid (0.5 mL) at room temperature. The resulting mixture was
stirred at 90.degree. C. for 3 hours. Then it was evaporated to
remove excess triethoxymethane and the crude product is achieved.
The crude product was diluted with ethyl acetate (50 mL), washed
with aqueous sodium bicarbonate and saturated brine successively.
The organic layer was dried over anhydrous sodium sulfate and
filtered. The filtrate was concentrated to give
5-bromo-6-methoxy-1H-benzo[d]imidazole 60d (430.0 mg, 1.9 mmol,
brown solid) as the desired product. Yield: 95%.
[0580] MS m/z (ESI): 227 & 229 [M+1].
[0581] Step 4
1-tert-Butoxycarbonyl-5-bromo-6-methoxy-benzoimidazole
3-tert-Butoxycarbonyl-5-bromo-6-methoxy-benzoimidazole
[0582] To the mixture of 5-bromo-6-methoxy-1H-benzoimidazole 60d
(430.0 mg, 1.9 mmol), triethylamine (383.0 mg, 3.8 mmol),
4-dimethylaminopyridine (23.0 mg, 0.2 mmol) and tetrahydrofuran
(15.0 mL) was added di-t-butyl dicarbonate (585.0 mg, 2.9 mmol) at
room temperature. The resulting mixture was stirred at 45.degree.
C. for 12 hours. Then it was diluted with dichloromethane (100.0
mL) and washed with brine (20.0 mL.times.3). The organic layer was
dried over anhydrous sodium sulfate and filtered. The filtrate was
concentrated to give a mixture of
1-tert-butoxycarbonyl-5-bromo-6-methoxy-benzoimidazole 60e and
3-tert-butoxycarbonyl-5-bromo-6-methoxy-benzoimidazole 61e (450.0
mg, 1.4 mmol, white solid). Yield: 73%.
[0583] MS m/z (ESI): 327 & 329 [M+1];
[0584] Step 5
1-tert-Butoxycarbonyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxy-
benzimidazole
3-tert-Butoxycarbonyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxy-
benzimidazole
[0585] To the mixture of
1-tert-butoxycarbonyl-5-bromo-6-methoxy-benzoimidazole 60e and
3-tert-butoxycarbonyl-5-bromo-6-methoxy-benzoimidazole 61e (200.0
mg, 0.61 mmol), N-(6-aminopyrimidin-4-yl)cyclopropanecarboxamide
(145.0 mg, 0.81 mmol), cesium carbonate (480.0 mg, 1.47 mmol) and
1,4-dioxane (2 mL) were added
tris(dibenzylideneacetone)dipalladium(0) (7.0 mg, 0.07 mmol) and
dicyclohexyl(2',4',6'-triisopropyl-3,6-dimethoxy-[1,1'-biphenyl]-2-yl-
)phosphine (79.0 mg, 0.15 mmol) at room temperature under argon.
Then the resulting mixture was stirred at 100.degree. C. for 1.5
hours under argon. It was cooled to room temperature, diluted with
dichloromethane (20 mL) and filtered. The filtrate was concentrated
under reduced pressure. Purification of the residue using Prep TLC
(dichloromethane/methanol=25:1)
gavel-tert-butoxycarbonyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-me-
thoxybenzimidazole 60 (40.0 mg, 0.094 mmol, white solid) and
3-tert-butoxycarbonyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methox-
ybenzimidazole 61 (60.0 mg, 0.142 mmol, white solid). Yield:
38%.
1-tert-butoxycarbonyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxy-
benzimidazole 60
[0586] MS m/z (ESI): 425 [M+1],
[0587] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.72 (s, 1H), 8.68
(brs, 1H), 8.45 (s, 1H), 8.31 (s, 1H), 7.58 (s, 2H), 7.36 (s, 1H),
3.94 (s, 3H), 1.72 (s, 9H), 1.60-1.55 (m, 1H), 1.11-1.07 (m, 2H),
0.94-0.87 (m, 2H).
3-tert-butoxycarbonyl-5-(6-cyclopropionamidopyrimidin-4-ylamino)-6-methoxy-
benzimidazole 61
[0588] MS m/z (ESI): 425 [M+1],
[0589] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.91 (s, 1H), 8.95
(s, 1H), 8.46 (s, 1H), 8.42 (s, 1H), 7.71 (s, 1H), 7.65 (s, 1H),
7.28 (s, 1H), 3.98 (s, 3H), 1.71 (s, 9H), 1.69-1.64 (m, 1H),
1.14-1.10 (m, 2H), 0.94-0.88 (m, 2H).
Example 62
5-(5-Methylaminopyrimidin-4-yl) amino)-6-methoxy-1H-indazole
formate
##STR00094##
[0591] The synthetic procedures were similar to that in Example 55.
Using 4-chloro-5-methylaminopyrimidine instead of
4-chloro-6-methoxypyrimidine. The desired product:
5-(5-methylaminopyrimidin-4-yl) amino)-6-methoxy-1H-indazole
formate (1.4 mg, 0.005 mmol, white solid) was obtained. Yield:
25%.
[0592] MS m/z (ESI): 271 [M+1],
[0593] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.97 (s, 1H), 9.13
(s, 1H), 8.20 (s, 1H), 7.98 (s, 1H), 7.95 (s, 1H), 7.56 (s, 1H),
7.08 (s, 1H), 6.28 (s, 1H), 3.86 (s, 3H), 2.82 (d, J=3.6 Hz,
3H).
Example 63
N-(6-((6-Methoxypyrazolo[1,5-a]pyridin-5-yl)amino)pyrimidin-4-yl)cycloprop-
anecarboxamide
##STR00095##
[0595] Step 1
3-Methoxypyridine 1-oxide
[0596] To the solution of 3-methoxypyridine 63a (1.1 g, 10.0 mmol)
in acetic acid (20 mL) was added hydrogen peroxide (2.0 mL, 20.0
mmol, 30% aqueous solution) at room temperature. Then the resulting
mixture was stirred at 80.degree. C. for 5 hours. The mixture was
concentrated, and 4 M aqueous sodium hydroxide was added until pH
>8. It was extracted with dichloromethane (10 mL.times.3). The
combined organic layer was washed with water (100 mL), dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated to give pure 3-methoxypyridine N-oxide 63b (0.42 g,
3.4 mmol, white solid). Yield: 34%. MS m/z (ESI): 26 [M+1].
[0597] Step 2
3-Methoxy-4-nitropyridine N-oxide
[0598] 3-Methoxypyridine N-oxide 63b (0.42 g, 3.4 mmol) was
dissolved in concentrated sulfuric acid (1.25 mL, 98%),
concentrated nitric acid (1.0 mL, 68%) was added slowly with
stirring in an ice bath. Then the resulting mixture was stirred at
85.degree. C. for 6 hours. The mixture was poured into ice water, 4
M aqueous sodium hydroxide was added to adjust the pH until pH
>7. The mixture was extracted with ethyl acetate (3.times.10
mL). The combined organic layer was washed with water (100 mL),
dried over anhydrous sodium sulfate and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using flash column chromatography
(dichloromethane/methanol=1:0-10:1) gave desired
3-methoxy-4-nitropyridine N-oxide 63c (0.37 g, 2.2 mmol, yellow
solid). Yield: 65%.
[0599] MS m/z (ESI): 172 [M+1],
[0600] Step 3
3-Methoxy-4-amino pyridine
[0601] To the solution of 3-methoxy-4-nitropyridine 1-oxide 63c
(0.37 g, 2.2 mmol) in methanol (100 mL) was added Raney nickle (100
mg). The resulting mixture was stirred at room temperature for 1.5
hours under hydrogen atmosphere. Then it was filtered through
celite to remove Raney nickle. The filtrate was concentrated to
give 3-methoxy-4-amino pyridine 63d (0.14 g, 1.1 mmol, pale yellow
liquid) as a crude product. Yield: 50%.
[0602] MS m/z (ESI): 125 [M+1],
[0603] Step 4
1-tert-Butoxycarbonyl-3-methoxy-4-aminopyridine
[0604] To the solution of 3-methoxy-4-amino pyridine 63d (0.14 g,
1.1 mmol) in tetrahydrofuran (10 mL) were added
N,N-diisopropylethylamine (173 mg, 1.3 mmol) and di-t-butyl
dicarbonate (366 mg, 1.7 mmol) in an ice bath. Then the resulting
mixture was stirred at room temperature overnight. It was
concentrated under reduced pressure. Purification of the residue
using flash column chromatography
(dichloromethane/methanol=1:0-10:1) gave
1-tert-butoxycarbonyl-3-methoxy-4-aminopyridine 63e (0.22 g, 1.0
mmol, white solid).
[0605] Yield: 91%.
[0606] MS m/z (ESI): 225 [M+1],
[0607] Step 5
1-Amino-4-((t-butoxycarbonyl)amino)-3-methoxypyridin-1-ium
2,4-dinitrophenolate
[0608] To the solution of
1-tert-butoxycarbonyl-3-methoxy-4-aminopyridine 63e (0.22 g, 1.0
mmol) in acetonitrile (30 mL) was added
O-(2,4-dinitrophenyl)hydroxylamine (225 mg, 1.1 mmol). Then the
resulting mixture was stirred at 40.degree. C. for overnight. It
was concentrated to give
1-amino-4-((t-butoxycarbonyl)amino)-3-methoxypyridin-1-ium
2,4-dinitrophenolate 63f (0.45 g, 1.0 mmol, light yellow liquid).
Yield: 100%.
[0609] MS m/z (ESI): 240 [M+1];
[0610] Step 6
Ethyl
5-((t-butoxycarbonyl)amino)-6-methoxypyrazolo[1,5-a]pyridine-3-carbo-
xylate
[0611] To a solution of
1-amino-4-((t-butoxycarbonyl)amino)-3-methoxypyridin-1-ium
2,4-dinitrophenolate 63f (0.45 g, 1.0 mmol) and ethyl propiolate
(108 mg, 1.1 mmol) in N,N-dimethylformamide (10 mL) in an ice bath
was added potassium carbonate (193 mg, 1.4 mmol). The resulting
mixture was stirred at room temperature overnight. Then it was
quenched with water and extracted with ethyl acetate (10
mL.times.3). The organic layer was dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated under reduced
pressure. Purification of the residue using flash column
chromatography (petroleum ether/ethyl acetate=1:0-5:1) gave ethyl
5-((t-butoxycarbonyl)amino)-6-methoxypyrazolo[1,5-a]pyridine-3-carboxylat-
e 63g (0.15 g, 0.45 mmol, yellow solid). Yield: 40%.
[0612] MS m/z (ESI): 336 [M+1],
[0613] Step 7
6-Methoxypyrazolo[1,5-a]pyridin-5-amine
[0614] The solution of ethyl
5-((t-butoxycarbonyl)amino)-6-methoxypyrazolo[1,5-a]pyridine-3-carboxylat-
e 63g (0.15 g, 0.45 mmol) in concentrated sulfuric acid (2 mL, 98%)
and water (2 mL) was stirred at 100.degree. C. for 4 hours. Then 6
M aqueous sodium hydroxide was added until pH >7. Then it was
extracted with dichloromethane (10 mL.times.3). The combined
organic layer was dried over anhydrous sodium sulfate and filtered.
The filtrate was concentrated to give
6-methoxypyrazolo[1,5-a]pyridin-5-amine 63h (72 mg, 0.44 mmol,
light yellow solid) as a crude product. Yield: 99%.
[0615] MS m/z (ESI): 164 [M+1],
[0616] Step 8
N-(6-((6-Methoxypyrazolo[1,5-a]pyridin-5-yl)amino)pyrimidin-4-yl)cycloprop-
anecarboxamide
[0617] To the mixture of
N-(6-chloropyrimidine-4-yl)cyclopropylcarboxamide 63h (5 mg, 0.03
mmol) 6-methoxypyrazolo[1,5-a]pyridin-5-amine (6 mg, 0.03 mmol),
and cesium carbonate (30 mg, 0.09 mmol) in 1,4-dioxane (1 mL) were
added tris(dibenzylideneacetone)dipalladium(0) (3.2 mg, 0.003 mmol)
and
dicyclohexyl(2',4',6'-triisopropyl-3,6-dimethoxy-[1,1'-biphenyl]-2-yl)pho-
sphine (3.6 mg, 0.006 mmol) at room temperature under argon. Then
the resulting mixture was stirred at 110.degree. C. for 1 hour
under argon in an oil bath. It was cooled to room temperature,
diluted with methanol (5.0 mL) and filtered. The filtrate was
concentrated to give crude product which was purified with Prep
HPLC (water (with 0.8% ammonia bicarbonate), 10%.about.40%
acetonitrile, 15 minutes) to give
N-(6-((6-methoxypyrazolo[1,5-a]pyridin-5-yl)amino)pyrimidin-4-yl)cyclopro-
panecarboxamide 63 (2.0 mg, 0.006 mmol, white solid). Yield:
20%.
[0618] MS m/z (ESI): 325 [M+1];
[0619] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.83 (s, 1H), 9.23
(s, 1H), 8.43 (d, J=7.6 Hz, 1H), 8.34 (s, 1H), 7.94 (s, 1H), 7.61
(s, 1H), 7.41 (d, J=7.6 Hz, 1H), 6.66 (s, 1H), 3.90 (s, 3H),
2.02-2.00 (m, 1H), 0.85-0.75 (m, 4H).
Example 64
6-Methoxy-N-(4-{1-methyl-7-[(3S)-3-methylpiperazin-1-yl]-1H-benzimidazol-2-
-yl} pyrimidine-5-)-1H-indazol-5-amine
##STR00096## ##STR00097##
[0621] Step 1
1-tert-Butoxycarbonyl
(2S)-4-(2-fluoro-3-nitrophenyl)-2-methylpiperazine
[0622] To the mixture of 1-bromo-2-fluoro-3-nitrobenzene 64a (3.1
g, 14.0 mmol), 1-tert-butoxycarbonyl (2S)-2-methylpiperazine (4.2
g, 21.0 mmol) and 1,4-dioxane (50 mL) were added
tris(dibenzylideneacetone)dipalladium(0) (0.6 g, 0.7 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.8 g, 1.4 mmol)
and cesium carbonate (9.1 g, 28 mmol) at room temperature under
argon. The resulting mixture was stirred at 110.degree. C. for 16
hours under argon. Then it was cooled to room temperature and
extracted with dichloromethane (100 mL.times.3). The combined
organic layer was washed with brine (100 mL), dried over anhydrous
sodium sulfate and filtered. The filtrate was concentrated and the
residue was purified by flash column chromatography (petroleum
ether/ethyl acetate=5:1) to give 1-tert-butoxycarbonyl
(2S)-4-(2-fluoro-3-nitrophenyl)-2-methylpiperazine 64b (2.6 g, 7.7
mmol, red oil). Yield: 55%.
[0623] MS m/z (ESI): 340 [M+1].
[0624] Step 2
1-tert-Butoxycarbonyl
(2S)-2-methyl-4-[2-(methylamino)-3-nitrophenyl]piperazine
[0625] 1-tert-Butoxycarbonyl
(2S)-4-(2-fluoro-3-nitrophenyl)-2-methylpiperazine 64b (2.4 g, 7.0
mmol) was mixed with methanamine (2 M in tetrahydrofuran, 15 mL)
and N,N-dimethylformamide (20 mL), the resultant mixture was
stirred at 130.degree. C. for 16 hours. Then it was cooled to room
temperature and extracted with dichloromethane (100 mL.times.3).
The combined organic layer was washed with brine (100 mL), dried
over anhydrous sodium sulfate and filtered. The filtrate was
concentrated and the residue was purified by flash column
chromatography (n-hexane/ethyl acetate=5:1) to give
1-tert-butoxycarbonyl
(2S)-2-methyl-4-[2-(methylamino)-3-nitrophenyl] piperazine 64c (1.9
g, 5.4 mmol, red oil). Yield: 78%.
[0626] MS m/z (ESI): 351 [M+1],
[0627] Step 3
1-tert-Butoxycarbonyl
(2S)-2-methyl-4-[2-(methylamino)-3-aminophenyl]piperazine
[0628] 1-tert-Butoxycarbonyl
(2S)-2-methyl-4-[2-(methylamino)-3-nitrophenyl]piperazine 64c (0.7
g, 2.0 mmol), Pd/C (containing 55% of water, 0.1 g) and methanol
(20 mL) were mixed and stirred at room temperature for 2 hours
under hydrogen atmosphere. Then it was filtered. The filtrate was
concentrated under reduced pressure and the residue was purified by
flash column chromatography (petroleum ether/ethyl acetate=2:1).
1-tert-butoxycarbonyl
(2S)-2-methyl-4-[2-(methylamino)-3-aminophenyl] piperazine 64d (0.4
g, 1.2 mmol, yellow oil) was obtained. Yield: 60%.
[0629] MS m/z (ESI): 321 [M+1],
[0630] Step 4
1-tert-Butoxycarbonyl
(2S)-4-[2-(4-aminopyrimidin-5-yl)-1-methyl-1H-benzimidazol-7-yl]-2-methyl-
piperazine
[0631] 1-tert-Butoxycarbonyl
(2S)-2-methyl-4-[2-(methylamino)-3-aminophenyl]piperazine 64d
(160.0 mg, 0.5 mmol), 2-(7-benzotriazole)-N, N,
N',N'-tetramethylurea hexafluorophosphate (266.0 mg, 0.7 mmol),
N,N-diisopropylethylamine (194.0 mg, 1.5 mmol) and
N,N-dimethylformamide (10 mL) were mixed and stirred at room
temperature for 10 minutes at room temperature. Then
4-aminopyrimidine-5-methanoic acid (84.0 mg, 0.6 mmol) was added to
the above mixture. The resulting mixture was stirred at room
temperature for 2 hours. Then it was diluted with water and
extracted with dichloromethane (50 mL.times.3). The combined
organic layer was washed with brine (100 mL), dried over anhydrous
sodium sulfate and filtered. The filtrate was concentrated and the
residue was dissolved in 10 ml of acetic acid directly and reacted
for 1 hour at 130.degree. C., cooled to room temperature,
concentrated under reduced pressure and diluted with
dichloromethane. The pH of the resultant mixture is adjusted to
pH=8.about.10 by addition of saturated sodium bicarbonate aqueous
solution, further extracted with dichloromethane (50 mL.times.3).
The combined organic layer was washed with brine (100 mL), dried
over anhydrous sodium sulfate and filtered. The filtrate was
concentrated and the residue was purified by flash column
chromatography (petroleum ether/ethyl acetate=2:1), and
1-tert-butoxycarbonyl
(2S)-4-[2-(4-aminopyrimidin-5-yl)-1-methyl-1H-benzimidazol-7-yl]-2-methyl-
piperazine 64e (85.0 mg, 0.2 mmol, yellow solid) was obtained.
Yield: 40%.
[0632] MS m/z (ESI): 424 [M+1],
[0633] Step 5
1-tert-Butoxycarbonyl-5-[(5-{7-[(3S)-4-(tert-butoxycarbonyl)-3-methylpiper-
azin-1-yl]-1-methyl-1H-Benzimidazol-2-yl} pyrimidin-4-yl)
amino]-6-methoxyindazole
[0634] To the mixture of 1-tert-butoxycarbonyl
(2S)-4-[2-(4-aminopyrimidin-5-yl)-1-methyl-1H-benzimidazol-7-yl]-2-methyl-
piperazine 64e (42.0 mg, 0.1 mmol), t-butyl
5-bromo-6-methoxy-1H-indazole-1-carboxylate (32.0 mg, 0.10 mmol)
and 1,4-dioxane (2 mL) were added
tris(dibenzylideneacetone)dipalladium(0) (9.0 mg, 0.01 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (12.0 mg, 0.02
mmol) and cesium carbonate (98 mg, 0.3 mmol) at room temperature
under argon. The resulting mixture was stirred at 120.degree. C.
for 1 hour with argon under microwave conditions. Then it was
cooled to room temperature, diluted with dichloromethane (10 mL)
and filtered. The filtrate was concentrated and the residue was
purified by Prep TLC (petroleum ether/ethyl acetate=1:3).
1-tert-butoxycarbonyl-5-[(5-{7-[(3S)-4-(tert-butoxycarbonyl)-3-methylpipe-
razin-1-yl]-1-methyl-1H-Benzimidazol-2-yl} pyrimidin-4-yl)
amino]-6-methoxyindazole 64f (14.0 mg, 0.02 mmol, yellow solid) was
obtained. Yield: 20%.
[0635] MS m/z (ESI): 670 [M+1],
[0636] Step 6
6-Methoxy-N-(4-{1-methyl-7-[(3S)-3-methylpiperazin-1-yl]-1H-benzimidazol-2-
-yl} pyrimidine-5-)-1H-indazol-5-amine
[0637]
1-tert-Butoxycarbonyl-5-[(5-{7-[(3S)-4-(tert-butoxycarbonyl)-3-meth-
ylpiperazin-1-yl]-1-methyl-1H-Benzimidazol-2-yl} pyrimidin-4-yl)
amino]-6-methoxyindazole 64f (9.0 mg, 0.015 mmol), dichloromethane
(2 mL) and trifluoroacetic acid (1 mL) were mixed and stirred at
room temperature for 1 hour. Then it was concentrated and diluted
with dichloromethane. The pH value was adjusted to 8.about.10 with
saturated aqueous sodium bicarbonate solution. The organic layer
was separated and the aqueous layer was extracted with
dichloromethane (10 mL.times.3). The combined organic layer was
washed with brine (10 mL), dried over anhydrous sodium sulfate and
filtered. The filtrate was concentrated under reduced pressure.
Purification of the residue using Prep TLC
(dichloromethane/methanol=8:1) gave
6-methoxy-N-(4-{1-methyl-7-[(3S)-3-methylpiperazin-1-yl]-1H-benzimidazol--
2-yl} pyrimidine-5-)-1H-indazol-5-amine 64 (4.0 mg, 0.01 mmol,
yellow solid). Yield: 60%.
[0638] MS m/z (ESI): 470 [M+1],
[0639] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 11.18 (s, 1H),
8.99 (s, 1H), 8.77 (s, 1H), 8.56 (s, 1H), 7.96 (s, 1H), 7.53 (d,
J=8.0 Hz, 1H), 7.23 (d, J=8.0 Hz, 1H), 7.01 (d, J=8.0 Hz, 1H), 6.86
(s, 1H), 4.19 (s, 3H), 3.95 (s, 3H), 3.20-3.09 (m, 4H), 2.59-2.49
(m, 2H), 2.28-2.23 (m, 1H), 1.13 (d, J=6.0 Hz, 3H).
Example 65
N-(6-(1H-Imidazol-1-yl)pyrimidin-4-yl)-6-methoxy-1H-indazol-5-amine
formate
##STR00098##
[0640] Step 1
6-(1H-Imidazol-1-yl)pyrimidin-4-amine
[0641] 6-Chloropyrimidin-4-amine 65a (200 mg, 1.5 mmol), imidazole
(116 mg, 1.7 mmol), cesium carbonate (978 mg, 3.0 mmol) and
N,N-dimethylformamide (2 mL) were mixed and stirred at 120.degree.
C. for 8 hours. Then it was cooled to room temperature, diluted
with dichloromethane (10 mL) and filtered. The filtrate was
concentrated to give the crude product. The crude product was
washed with water (50 mL) and dried to give
6-(1H-imidazol-1-yl)pyrimidin-4-amine 65b (90 mg, 0.56 mmol, white
solid). Yield: 36%.
[0642] MS m/z (ESI): 162 [M+1],
[0643] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.45 (s, 1H), 8.33
(s, 1H), 7.82 (s, 1H), 7.20 (s, 2H), 7.12 (s, 1H), 6.57 (s,
1H).
[0644] Step 2
N-(6-(1H-Imidazol-1-yl)pyrimidin-4-yl)-6-methoxy-1H-indazol-5-amine
formate
[0645] To the mixture of 6-(1H-imidazol-1-yl)pyrimidin-4-amine 65b
(30.0 mg, 0.18 mmol), t-butyl
5-bromo-6-methoxy-1H-indazole-1-carboxylate (66.4 mg, 0.20 mmol)
and 1,4-dioxane (1 mL) were added
tris(dibenzylideneacetone)dipalladium(0) (16.5 mg, 0.018 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (10.5 mg, 0.018
mmol) and cesium carbonate (117 mg, 0.36 mmol) at room temperature
under argon. The resulting mixture was stirred at 140.degree. C.
for 1 hour with argon under microwave conditions. Then it was
cooled to room temperature, diluted with dichloromethane (10 mL)
and filtered. The filtrate was concentrated under reduced pressure.
Purification of the residue using Prep HPLC (water (with 0.2%
formic acid), 10%.about.30% acetonitrile, 15 minutes) gave
N-(6-(1H-imidazol-1-yl)pyrimidin-4-yl)-6-methoxy-1H-indazol-5-amine
formate salt 65 (1.2 mg, 0.004 mmol, white solid). Yield: 2%.
[0646] MS m/z (ESI): 308 [M+1],
[0647] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 12.85 (s, 1H), 9.11
(s, 1H), 8.62-8.41 (m, 2H), 8.21-8.04 (m, 2H), 7.97 (s, 1H), 7.85
(s, 1H), 7.17 (s, 1H), 7.07 (s, 1H), 6.92 (s, 1H), 3.89 (s,
3H).
Example 66
5-((6-Aminopyrimidin-4-yl)amino)-4-methoxyisoindolin-1-one
hydrochloride
##STR00099##
[0648] Step 1 3-Hydroxy-2-methyl-4-nitrobenzoic acid
[0649] To the solution of 3-hydroxy-2-methylbenzoic acid 66a (20 g,
131.6 mmol) in acetic acid (160 mL) was added nitric acid (19.6 mL,
70%) slowly at 0.degree. C. The mixture was stirred at room
temperature for 1 hour. Then it was poured into ice water (500 mL).
The precipitated yellow solid was filtered.
3-Hydroxy-2-methyl-4-nitrobenzoic acid 66b (10.4 g, 52.5 mmol,
yellow solid) was obtained. Yield: 40%.
[0650] MS m/z (ESI): 198 [M+1],
[0651] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 11.09 (s, 1H),
8.04 (d, J=9.0 Hz, 1H), 7.50 (d, J=9.0 Hz, 1H), 2.60 (s, 3H).
[0652] Step 2
Methyl 3-methoxy-2-methyl-4-nitrobenzoate
[0653] To the mixture of 3-hydroxy-2-methyl-4-nitrobenzoic acid 66b
(10.3 g, 52.0 mmol) with acetone (240 mL) were added dimethyl
sulfate (17.4 g, 140.5 mmol) and potassium carbonate (31 g, 234.0
mmol) at room temperature. The mixture was stirred at 60.degree. C.
for 2 hours. Then it was concentrated under reduced pressure.
Purification of the residue using flash column chromatography
(n-Hexane/ethyl acetate=1:0-10:1) gave methyl
3-methoxy-2-methyl-4-nitrobenzoate 66c (11.2 g, 50.0 mmol, yellow
solid). Yield: 94%.
[0654] MS m/z (ESI): 226 [M+1],
[0655] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.68 (d, J=8.5 Hz,
1H), 7.61 (d, J=8.5 Hz, 1H), 3.94 (s, 3H), 3.91 (s, 3H), 2.55 (s,
3H).
[0656] Step 3
Methyl 2-(bromomethyl)-3-methoxy-4-nitrobenzoate
[0657] To the solution of methyl 3-methoxy-2-methyl-4-nitrobenzoate
66c (5.6 g, 24.9 mmol) in acetonitrile (100 mL) were added
N-bromosuccinimide (5.4 g, 30.5 mmol) and azodiisobutyronitrile (82
mg, 0.5 mmol) at room temperature. The mixture was stirred at
80.degree. C. overnight. Then it was quenched with water (100 mL)
and extracted with ethyl acetate (50 mL.times.3). The combined
organic layer was washed with brine (50 mL.times.2), dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using flash column chromatography (n-hexane/ethyl
acetate=1:0.about.5:1) gave methyl
2-(bromomethyl)-3-methoxy-4-nitrobenzoate 66d (7.5 g, 24.8 mmol,
yellow oil). Yield: 99%.
[0658] MS m/z (ESI): 224 [M-Br]
[0659] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.83-7.75 (m, 2H),
5.05 (s, 2H), 4.05 (s, 3H), 3.99 (s, 3H).
[0660] Step 4
4-Methoxy-5-nitroisoindolin-1-one
[0661] To the solution of methyl
2-(bromomethyl)-3-methoxy-4-nitrobenzoate 66d (7.5 g, 24.8 mmol) in
methanol (80 mL) were added triethylamine (3.0 g, 30.0 mmol) and
ammonia (25 mL, 175 mmol, 7 M in methanol) at room temperature. The
mixture was stirred at 70.degree. C. for 4 hours. Then it was
concentrated under reduced pressure. After the residue was
recrystallized in methanol. 4-Methoxy-5-nitroisoindolin-1-one 66e
(3.8 g, 18.3 mmol, yellow solid) was obtained. Yield: 74%.
[0662] MS m/z (ESI): 209 [M+1],
[0663] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.07 (s, 1H), 7.92
(d, J=8.0 Hz, 1H), 7.46 (d, J=8.0 Hz, 1H), 4.75 (s, 2H), 4.08 (s,
3H).
[0664] Step 5
5-Amino-4-methoxyisoindolin-1-one
[0665] To the mixture of 4-methoxy-5-nitroisoindolin-1-one 66e (3.8
g, 18.3 mmol) in methanol (300 mL) was added Pd/C (1.0 g, 26 wt %,
55% moisture content) at room temperature. The mixture was stirred
at 50.degree. C. under hydrogen atmosphere for overnight. Then it
was filtered with celite to remove Pd/C. The filtrate was
concentrated under reduced pressure. Purification of the residue
using column chromatography (dichloromethane/methanol=1:0.about.
10:1) gave 5-amino-4-methoxyisoindolin-1-one 66f (3.2 g, 18.3 mmol,
light yellow solid). Yield: 98%.
[0666] MS m/z (ESI): 179 [M+1],
[0667] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.02 (s, 1H), 7.12
(t, J=8.0 Hz, 1H), 6.73 (d, J=8.0 Hz, 1H), 5.43 (s, 2H), 4.36 (s,
2H), 3.86-3.70 (m, 3H).
[0668] Step 6
5-((6-Aminopyrimidin-4-yl)amino)-4-methoxyisoindolin-1-one
hydrochloride
[0669] To the solution of 5-amino-4-methoxyisoindolin-1-one 66f
(0.13 g, 0.73 mmol) in dioxane (10 mL) was added a solution of
hydrochlorie in methanol (3.5 mL, 1 M, 3.5 mmol). The reaction tube
was sealed, and the mixture was reacted at 130.degree. C. for
overnight. Then it was concentrated under reduced pressure.
Purification of the residue using Prep-HPLC (water (0.2% formic
acid), 10%.about.30% acetonitrile, 15 minutes) gave
5-((6-aminopyrimidin-4-yl)amino)-4-methoxyisoindolin-1-one
hydrochloride salt 66 (5 mg, 0.018 mmol, white solid). Yield:
2.5%.
[0670] MS m/z (ESI): 272 [M+1],
[0671] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.45 (s, 1H), 8.38
(s, 1H), 8.09 (d, J=8.2 Hz, 1H), 8.05 (s, 1H), 7.31 (d, J=8.2 Hz,
1H), 6.39 (s, 2H), 5.92 (s, 1H), 4.52 (s, 2H), 3.88 (s, 3H).
Example 67
5-((6-Cyclopropanamidopyrimidin-4-yl)
amino)-4-methoxyisodihydroindole-1-one
##STR00100##
[0673] Step 1
N-(6-Chloropyrimidin-4-yl)cyclopropanecarboxamide
[0674] To the solution of 6-chloropyrimidin-4-amine 67a (129 mg,
1.0 mmol) in tetrahydrofuran (5 mL) were added cyclopropanecarbonyl
chloride (208 mg, 2.0 mmol) and potassium carbonate (414 mg, 3.0
mmol) at room temperature. The mixture was stirred at 70.degree. C.
for overnight. Then it was concentrated under reduced pressure.
Purification of the residue using column chromatography (petroleum
ether/ethyl acetate=1:0-1:1) gave
N-(6-chloropyrimidin-4-yl)cyclopropanecarboxamide 67b (88.6 mg, 0.3
mmol, white solid). Yield: 45%.
[0675] MS m/z (ESI): 198 & 200 [M+1],
[0676] Step 2
5-((6-Cyclopropanamidopyrimidin-4-yl)
amino)-4-methoxyisodihydroindole-1-one
[0677] To the solution of
N-(6-chloropyrimidin-4-yl)cyclopropanecarboxamide 67b (49 mg, 0.25
mmol) in acetic acid (5 mL) was added
5-amino-4-methoxyisoindolin-1-one (45 mg, 0.25 mmol) at room
temperature. The mixture was stirred at 110.degree. C. overnight.
Then it was concentrated under reduced pressure. Purification of
the residue using Prep-HPLC (water (0.2% formic acid),
10%.about.30% acetonitrile, 15 minutes) gave
5-((6-cyclopropanamidopyrimidin-4-yl)
amino)-4-methoxyisodihydroindole-1-one 67 (2.9 mg, 0.0085 mmol,
white solid). Yield: 3%.
[0678] MS m/z (ESI): 340 [M+1],
[0679] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.83 (s, 1H), 9.10
(s, 1H), 8.54 (s, 1H), 8.37 (s, 1H), 8.10 (d, J=8.1 Hz, 1H), 7.66
(s, 1H), 7.34 (d, J=8.1 Hz, 1H), 4.55 (s, 2H), 3.89 (s, 3H),
2.06-2.02 (m, 1H), 0.88-0.80 (m, 4H).
Example 68
5-((6-(Pyridin-2-ylamino) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindol-1-one
##STR00101##
[0681] Step 1
6-Chloro-N-(pyridin-2-yl)pyrimidin-4-amine
[0682] 6-Chloropyrimidin-4-amine 68a (129 mg, 1.0 mmol),
2-bromopyridine (17.2 mg, 1.1 mmol), cesium carbonate (978 mg, 3
mmol), tris(dibenzylideneacetone)-dipalladium(0) (92 mg, 0.1 mmol),
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (116 mg, 0.2 mmol)
and dioxane (5 mL) were mixed and stirred at 110.degree. C. under
argon for 1 hour. Then it was filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using column chromatography (petroleum ether/ethyl acetate=0:1-1:1)
gave 6-chloro-N-(pyridin-2-yl)pyrimidin-4-amine 68b (116 mg, 0.56
mmol, white solid). Yield: 56%.
[0683] MS m/z (ESI): 207 & 209 [M+1],
[0684] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.57 (s, 1H), 8.37
(d, J=4.7 Hz, 1H), 7.96 (s, 1H), 7.77-7.58 (m, 2H), 7.29 (d, J=8.4
Hz, 1H), 7.06-6.99 (m, 1H).
[0685] Step 2
5-((6-(Pyridin-2-ylamino) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindol-1-one
[0686] To the mixture of 6-chloro-N-(pyridin-2-yl)pyrimidin-4-amine
68b (65 mg, 0.5 mmol) in butan-1-ol (2 mL) were added
5-amino-4-methoxyisoindolin-1-one (90 mg, 0.5 mmol) and a solution
of hydrochloric acid in methanol (2 mL, 4 M). The reaction tube was
sealed, and the mixture was stirred at 130.degree. C. overnight.
Then it was concentrated under reduced pressure. Purification of
the residue using Prep-HPLC (water (0.2% formic acid),
10%.about.30% acetonitrile, 15 minutes) gave
5-((6-(pyridin-2-ylamino) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindol-1-one 68 (4.4 mg, 0.0126 mmol,
white solid). Yield: 3%.
[0687] MS m/z (ESI): 349 [M+1],
[0688] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.83 (s, 1H), 8.87
(s, 1H), 8.50 (s, 1H), 8.31 (s, 1H), 8.27 (d, J=5.2 Hz, 1H), 8.10
(d, J=8.1 Hz, 1H), 7.69 (t, J=7.8 Hz, 1H), 7.48 (d, J=7.8 Hz, 2H),
7.34 (d, J=8.1 Hz, 1H), 6.94 (t, J=5.2 Hz, 1H), 4.56 (s, 2H), 3.91
(s, 3H).
Example 69
5-((6-(Pyrimidin-2-ylamino) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindol-1-one
##STR00102##
[0690] Step 1
N-(Pyrimidin-2-yl)pyrimidine-4,6-diamine
[0691] To the mixture of pyrimidine-4,6-diamine 69a (110.0 mg, 1.0
mmol) and sodium hydride (60% dispersion in mineral oil, 120.0 mg,
5.0 mmol) in N, N-dimethylacetamide (10.0 mL) was added
2-chloropyrimidine (114.0 mg, 1.0 mmol) at room temperature. The
mixture was stirred at 70.degree. C. for 2 hours. Then it was
cooled to room temperature and quenched with saturated aqueous
sodium bicarbonate (10 mL) and extracted with dichloromethane (30
mL.times.3). The combined organic layer was washed with brine (30
mL), dried over anhydrous sodium sulfate and then filtered. The
filtrate was concentrated under reduced pressure. Purification of
the residue using column chromatography
(dichloromethane/methanol=10:1) gave
N-(Pyrimidin-2-yl)pyrimidine-4,6-diamine 69b (47.0 mg, 0.20 mmol,
white solid). Yield: 25%.
[0692] MS m/z (ESI): 189 [M+1],
[0693] Step 2
5-((6-(Pyrimidin-2-ylamino) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindol-1-one
[0694] To the mixture of N-(Pyrimidin-2-yl)pyrimidine-4,6-diamine
69b (19.0 mg, 0.10 mmol), 5-bromo-4-methoxyisoindolin-1-one (25.0
mg, 0.10 mmol) and 1,4-dioxane (2.0 mL) were added
tris(dibenzylideneacetone)dipalladium(0) (9.0 mg, 0.01 mmol) and
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (12.0 mg, 0.02
mmol) at room temperature under argon. The mixture was stirred at
110.degree. C. for 1 hour in microwave oven with argen. Then it was
diluted with dichloromethane (10 mL) and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep-TLC (dichloromethane/methanol=20:1) gave
5-((6-(pyrimidin-2-ylamino) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindol-1-one 69 (7.0 mg, 0.02 mmol, white
solid). Yield: 20%.
[0695] MS m/z (ESI): 350 [M+1],
[0696] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.08 (s, 1H,), 9.03
(s, 1H,), 8.63 (d, J=4.4 Hz, 2H), 8.53 (s, 1H), 8.36 (s, 1H), 8.16
(d, J=8.0 Hz, 1H), 7.91 (s, 1H), 7.35 (d, J=8.0 Hz, 1H), 7.05 (t,
J=4.4 Hz, 1H), 4.58 (s, 2H), 3.93 (s, 3H).
Example 70
5-((6-(Pyrimidin-4-ylamino) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindol-1-one
##STR00103##
[0698] The synthetic procedure is similar to that of example 69.
Using 4-chloropyrimidine instead of 2-chloropyrimidine. The desired
compound 5-((6-(pyrimidin-4-ylamino) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindol-1-one (7 mg, 0.01 mmol, white
solid) was obtained. Yield: 20%.
[0699] MS m/z (ESI): 350 [M+1],
[0700] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.32 (s, 1H), 9.10
(s, 1H), 8.77 (s, 1H), 8.54 (s, 1H), 8.46 (d, J=6.0 Hz, 1H), 8.38
(s, 1H), 8.06 (d, J=8.0 Hz, 1H), 7.59 (d, J=6.0 Hz, 1H), 7.49 (s,
1H), 7.34 (d, J=8.0 Hz, 1H), 4.57 (s, 2H), 3.92 (s, 3H).
Example 71
5-((6-Amino-5-methoxypyrimidin-4-yl)amino)-4-methoxyisoindolin-1-one
##STR00104##
[0702] To the mixture of 6-chloro-5-methoxy-4-aminopyrimidine 71a
(16.0 mg, 0.1 mmol), 5-amino-4-methoxyisoindolin-1-one (18.0 mg,
0.1 mmol) and dioxane (1 mL) was added a solution of hydrochloric
acid in dioxane (0.1 mL, 4 M) at room temperature. The mixture was
stirred at 100.degree. C. under argon for 16 hours. Then it was
quenched with saturated aqueous sodium bicarbonate (10 mL). The
organic layer was separated and the aqueous layer was extracted
with dichloromethane (15 mL.times.2). The combined organic layer
was washed with brine (50 mL.times.2), dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated under reduced
pressure. Purification of the residue using Prep-TLC
(dichloromethane/methanol=10:1) gave
5-((6-amino-5-methoxypyrimidin-4-yl)amino)-4-methoxyisoindolin-1-one
71 (3 mg, 0.01 mmol, yellow solid). Yield: 10%.
[0703] MS m/z (ESI): 302 [M+1],
[0704] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.58 (d, J=8.4 Hz,
1H), 8.46 (s, 1H), 7.93 (s, 1H), 7.83 (s, 1H), 7.35 (d, J=8.4 Hz,
1H), 6.58 (s, 2H), 4.59 (s, 2H), 4.01 (s, 3H), 3.71 (s, 3H).
Example 72
5-((6-(Cyclopropionamido) pyrimidin-4-yl)
amino)-4-methoxy-2-methylisodihydroindol-1-one
##STR00105##
[0706] Step 1
4-Methoxy-2-methyl-5-nitroisoindolin-1-one
[0707] To the mixture of methyl
2-(bromomethyl)-3-methoxy-4-nitrobenzoate 72a (0.3 g, 1.0 mmol) in
methanol (10 mL) was added methylamine (1 mL, 10 M in water) at
room temperature. The mixture was stirred at room temperature for 1
hour. Then it was concentrated under reduced pressure. The residue
was recrystallized in methanol.
4-Methoxy-2-methyl-5-nitroisoindolin-1-one 72b (0.2 g, 0.9 mmol,
yellow solid) was obtained. Yield: 90%.
[0708] MS m/z (ESI): 223 [M+1].
[0709] Step 2
5-Amino-4-methoxy-2-methylisoindolin-1-one
[0710] To the mixture of 4-methoxy-2-methyl-5-nitroisoindolin-1-one
72b (0.2 g, 0.9 mmol) in methanol (50 mL) was added Pd/C (50 mg,
26% wt, 55% moisture content) at room temperature. The mixture was
stirred at 40.degree. C. under hydrogen for 12 hours. Then it was
filtered with celite to remove Pd/C. The filtrate was concentrated
under reduced pressure. Purification of the residue using column
chromatography (dichloromethane/methanol=1:0-10:1) gave
5-amino-4-methoxy-2-methylisoindolin-1-one 72c (0.15 g, 0.78 mmol,
light yellow solid).
[0711] Yield: 87%.
[0712] MS m/z (ESI): 193 [M+1],
[0713] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 7.09 (d, J=7.8 Hz,
1H), 6.72 (d, J=7.8 Hz, 1H), 5.47 (s, 2H), 4.47 (s, 2H), 3.80 (s,
3H), 3.00 (s, 3H).
Step 3
5-((6-(Cyclopropionamido) pyrimidin-4-yl)
amino)-4-methoxy-2-methylisodihydroindol-1-one
[0714] To the mixture of 5-amino-4-methoxy-2-methylisoindolin-1-one
72c (10 mg, 0.05 mmol),
N-(6-chloropyrimidin-4-yl)cyclopropanecarboxamide (10 mg, 0.05
mmol) and cesium carbonate (50 mg, 0.15 mmol) in 1,4-dioxane (1.0
mL) were added tris(dibenzylideneacetone)dipalladium(0) (4.2 mg,
0.005 mmol) and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene
(5.6 mg, 0.01 mmol) at room temperature under argon. The mixture
was stirred at 110.degree. C. for 1 hour in an oil bath. Then it
was cooled to room temperature, diluted with methanol (5 mL) and
filtered. The filtrate was concentrated under reduced pressure.
Purification of the residue using Prep-HPLC (water (0.2% formic
acid), 20%.about.60% acetonitrile, 15 minutes) gave
5-((6-(cyclopropionamido) pyrimidin-4-yl)
amino)-4-methoxy-2-methylisodihydroindol-1-one 72 (5.0 mg, 0.014
mmol, white solid). Yield: 28%.
[0715] MS m/z (ESI): 354 [M+1],
[0716] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.82 (s, 1H), 9.09
(s, 1H), 8.38 (s, 1H), 8.10 (d, J=8.1 Hz, 1H), 7.66 (s, 1H), 7.34
(d, J=8.1 Hz, 1H), 4.72-4.66 (m, 2H), 3.90 (s, 3H), 3.10-3.06 (m,
3H), 2.05-1.98 (m, 1H), 0.96-0.78 (m, 4H).
Example 73
5-((6-(Cyclopropionamido) pyrimidin-4-yl)
amino)-4-methoxy-2,3,3-trimethylisoindole-1-one
##STR00106##
[0718] Step 1
5-Bromo-4-methoxy-2,3,3-trimethylisoindolin-1-one
[0719] To the solution of 5-bromo-4-methoxyisoindolin-1-one 73a
(30.0 mg, 0.12 mmol) in N,N-dimethylformamide (10.0 mL) was added
sodium hydride (60% dispersion in mineral oil, 40.0 mg, 0.96 mmol)
in several portions at 0.degree. C. The mixture was stirred at room
temperature for 15 minutes. Then iodomethane (106.0 mg, 0.72 mmol)
was added dropwise and the resulting mixture was stirred at
60.degree. C. for 1 hour. Then it was cooled to room temperature,
quenched with water (30 mL) and extracted with dichloromethane (15
mL.times.3). The combined organic layer was washed with brine (30
mL), dried over anhydrous sodium sulfate and then filtered. The
filtrate was concentrated under reduced pressure. Purification of
the residue using Prep-TLC (dichloromethane/methanol=30:1) gave
5-bromo-4-methoxy-2,3,3-trimethylisoindolin-1-one 73b (20.0 mg,
0.071 mmol, yellow oil). Yield: 59%.
[0720] MS m/z (ESI): 284 & 286 [M+1],
[0721] Step 2
5-((6-(Cyclopropionamido) pyrimidin-4-yl)
amino)-4-methoxy-2,3,3-trimethylisoindole-1-one
[0722] To the mixture of
5-bromo-4-methoxy-2,3,3-trimethylisoindolin-1-one 73b (20.0 mg,
0.07 mmol), N-(6-aminopyrimidin-4-yl)cyclopropanecarboxamide (19.0
mg, 0.11 mmol) and cesium carbonate (92.6 mg, 0.28 mmol) in
1,4-dioxane (1.0 mL) were added
tris(dibenzylideneacetone)dipalladium(0) (6.1 mg, 0.007 mmol) and
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (7.8 mg, 0.014
mmol) at room temperature under argon. The mixture was stirred at
80.degree. C. for 3 hours. Then it was cooled to room temperature,
diluted with dichloromethane (10 mL) and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep-TLC (dichloromethane/methanol=15:1) gave
5-((6-(cyclopropionamido) pyrimidin-4-yl)
amino)-4-methoxy-2,3,3-trimethylisoindole-1-one 73 (10.0 mg, 0.026
mmol, white solid). Yield: 37%.
[0723] MS m/z (ESI): 382 [M+1],
[0724] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.82 (s, 1H), 9.31
(s, 1H), 8.34 (s, 1H), 7.80 (d, J=8.0 Hz, 1H), 7.59 (s, 1H), 7.39
(d, J=8.0 Hz, 1H), 3.74 (s, 3H), 2.90 (s, 3H), 2.09-1.91 (m, 1H),
1.49 (s, 6H), 7.65-7.63 (m, 4H).
Example 74
5-((5-Methoxypyrimidin-4-yl)
amino)-4-methoxyisodihydroindole-1-one
##STR00107##
[0726] 4-Chloro-5-methoxypyrimidine 74a (15.0 mg, 0.1 mmol),
5-amino-4-methoxyisoindolin-1-one (18.0 mg, 0.1 mmol) and acetic
acid (2 mL) were mixed and stirred at 70.degree. C. for 1 hour.
Then it was cooled to room temperature. White solid was formed and
filtered. The filter cake was washed with acetic acid (20 mL) and
dried to give 5-((5-methoxypyrimidin-4-yl)
amino)-4-methoxyisodihydroindole-1-one 74 (8.0 mg, 0.03 mmol, white
solid). Yield: 30%.
[0727] MS m/z (ESI): 287 [M+1],
[0728] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.93 (s, 1H), 8.74
(s, 1H), 8.59 (s, 1H), 8.24 (s, 1H), 7.80 (d, J=7.2 Hz, 1H), 7.41
(d, J=7.2 Hz, 1H), 4.66 (s, 2H), 4.05 (s, 3H), 3.94 (s, 3H).
Example 75
5-((5-(2-Methoxyethoxy) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindole-1-one
##STR00108##
[0729] Step 1
5-Bromo-4-methoxyisoindolin-1-one
[0730] To the solution of 5-amino-4-methoxyisoindolin-1-one 75a
(370.0 mg, 2.1 mmol) and copper(I) bromide (450.0 mg, 3.1 mmol) in
acetonitrile (10.0 mL) was added t-butyl nitrite (430.0 mg, 4.2
mmol) at room temperature. The resulting mixture was stirred at
50.degree. C. for 2 hours. Then it was cooled to room temperature
and quenched with diluted hydrochloric acid (5 mL, 1 M). The
mixture was diluted with dichloromethane (50 mL). The organic layer
was washed with brine (20 mL.times.3), dried over anhydrous sodium
sulfate and then filtered. The filtrate was concentrated under
reduced pressure. Purification of the residue using column
chromatography (dichloromethane/methanol=20/1) gave
5-bromo-4-methoxyisoindolin-1-one 75b (200 mg, 0.83 mmol, yellow
solid). Yield: 40%.
[0731] MS m/z (ESI): 242 & 244 [M+1],
[0732] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.68 (d, J=8.0 Hz,
1H), 7.48 (d, J=8.0 Hz, 1H), 7.10 (brs, 1H), 4.56 (s, 2H), 3.98 (s,
3H).
[0733] Step 2
5-((5-(2-Methoxyethoxy) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindole-1-one
[0734] To the mixture of 5-bromo-4-methoxyisoindolin-1-one 75b
(28.0 mg, 0.12 mmol), 5-(2-methoxyethoxy)-4-aminopyrimidine (20.0
mg, 0.12 mmol) and cesium carbonate (72.0 mg, 0.23 mmol) in
1,4-dioxane (2.0 mL) were added
tris(dibenzylideneacetone)dipalladium(0) (1.0 mg, 0.012 mmol) and
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (12.0 mg, 0.023
mmol) at room temperature under argon. The mixture was stirred at
120.degree. C. for 1 hour in microwave oven. It was cooled to room
temperature and filtered. The filtrate was concentrated.
Purification of the residue using Prep-HPLC (water (0.2% formic
acid), 0%.about.15% acetonitrile, 15 minutes) gave
5-((5-(2-methoxyethoxy) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindole-1-one 75 (4.0 mg, 0.012 mmol,
white solid). Yield: 10%.
[0735] MS m/z (ESI): 331 [M+1],
[0736] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.71 (d, J=8.2 Hz,
1H), 8.55 (s, 1H), 8.40 (s, 1H), 8.32 (s, 1H), 8.21 (brs, 1H), 7.41
(d, J=8.2 Hz, 1H), 4.63 (s, 2H), 4.35-4.33 (m, 2H), 4.03 (s, 3H),
3.78-3.74 (m, 2H), 3.39 (s, 3H).
Example 76
5-((5-(2-(4-morpholin) ethoxy) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindol-1-one formate
##STR00109##
[0738] The synthetic procedure is similar to that of example 75.
Using 4-(2-chloroethyl)morpholine instead of
1-bromine-2-methoxyethane. The desired compound
5-((5-(2-(4-morpholin) ethoxy) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindol-1-one formate was obtained. Yield:
25%.
[0739] MS m/z (ESI): 386 [M+1],
[0740] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.73 (d, J=8.0 Hz,
1H), 8.54 (s, 1H), 8.40 (s, 1H), 8.30 (s, 1H), 8.21 (s, 1H), 7.41
(d, J=8.0 Hz, 1H), 4.64 (s, 2H), 4.34-4.32 (m, 2H), 4.04 (s, 3H),
3.60-3.54 (m, 4H), 3.37-3.32 (m, 4H), 2.81-2.79 (m, 2H).
Example 77
5-((5-(2-(Dimethylamino) ethoxy) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindole-1-one dicarboxylate
##STR00110##
[0742] Step 1
5-(2-(Dimethylamino)ethoxy)-4-aminopyrimidine
[0743] 4-Amino-5-hydroxypyrimidine 77a (100.0 mg, 0.90 mmol),
anhydrous lithium hydroxide (65.0 mg, 2.70 mmol) and
N,N-dimethylformamide (3.0 mL) were mixed and stirred at room
temperature for 30 minutes. Then N,N-dimethyl-3-chloroethylamine
(125.0 mg, 0.90 mmol) was added to the above mixture. The resulting
mixture was stirred at 60.degree. C. for 5 hours. Then it was
cooled to room temperature and filtered. The filtrate was
concentrated under reduced pressure. Purification of the residue
using Prep-TLC (dichloromethane/methanol=10:1) gave
5-(2-(dimethylamino)ethoxy)-4-aminopyrimidine 77b (60.0 mg, 0.36
mmol, colorless oil). Yield: 40%.
[0744] MS m/z (ESI): 183 [M+1],
[0745] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.24 (s, 1H), 7.81
(s, 1H), 5.93 (brs, 2H), 4.23-4.18 (m, 2H), 3.08-3.04 (m, 2H), 2.60
(s, 6H).
[0746] Step 2
5-((5-(2-(Dimethylamino)ethoxy)pyrimidin-4-yl)amino)-4-methoxyisoindolin-1-
-one diformate
[0747] To the mixture of
5-(2-(Dimethylamino)ethoxy)-4-aminopyrimidine 77b (12.0 mg, 0.06
mmol), 5-bromo-4-methoxyisoindolin-1-one (10.0 mg, 0.04 mmol) and
cesium carbonate (40.0 mg, 0.12 mmol) in 1,4-dioxane (2.0 mL) were
added tris(dibenzylideneacetone)dipalladium(0) (4.0 mg, 0.004 mmol)
and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (4.0 mg, 0.008
mmol) at room temperature under argon. The mixture was stirred at
140.degree. C. for 1 hour in microwave oven. It was cooled to room
temperature and filtered. The filtrate was concentrated under
reduced pressure. Purification of the residue using Prep-HPLC
(water (0.2% formic acid), 0%-15% acetonitrile, 15 minutes) gave
5-((5-(2-(Dimethylamino)ethoxy)pyrimidin-4-yl)amino)-4-methoxyisoindolin--
1-one diformate 77 (2.0 mg, 0.006 mmol, white solid). Yield:
10%.
[0748] MS m/z (ESI): 344 [M+1],
[0749] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.70 (d, J=8.0 Hz,
1H), 8.55 (s, 1H), 8.42 (s, 1H), 8.40 (s, 1H), 8.26 (s, 1H), 8.21
(s, 1H), 7.96 (s, 1H), 7.42 (d, J=8.0 Hz, 1H), 4.64 (s, 2H), 4.30
(t, J=12.0 Hz, 2H), 2.90 (s, 3H), 2.78 (t, J=12.0 Hz, 2H), 2.76 (s,
6H).
Example 78
5-((5-(2-Hydroxyethoxy)pyrimidin-4-yl)amino)-4-methoxyisoindolin-1-one
##STR00111##
[0751] The synthetic procedure is similar to that of example 54.
Using 5-bromine-4-methoxyisoindolin-1-one instead of
1-t-butyloxoyl-5-amino-6-methoxy indazole. The desired compound:
5-((5-(2-Hydroxyethoxy)pyrimidin-4-yl)amino)-4-methoxyisoindolin-1-one
was obtained. Yield: 25%.
[0752] MS m/z (ESI): 317 [M+1],
[0753] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.53 (s, 1H), 8.51
(d, J=8.0, 1H), 8.41 (s, 1H), 8.35 (s, 1H), 8.18 (s, 1H), 7.41 (d,
J=8.0, 1H), 5.32-4.79 (brs, 1H), 4.62 (s, 2H), 4.22-4.21 (m, 2H),
3.80 (s, 3H), 3.82-3.79 (m, 2H).
Example 79
5-((7H-Pyrrolo[2,3-d]pyrimidin-4-yl)amino)-4-methoxyisoindolin-1-one
##STR00112##
[0755] The synthetic procedure is similar to that of example 74.
Using 4-chloropyrrolo[2,3-d]pyrimidine instead of
4-chloro-5-methoxypyrimidine. The desired compound:
5-((7H-Pyrrolo[2,3-d]pyrimidin-4-yl)amino)-4-methoxyisoindolin-1-one
(15.0 mg, 0.05 mmol, red solid) was obtained. Yield: 18%.
[0756] MS m/z (ESI): 296 [M+1],
[0757] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.80 (brs, 1H), 8.78
(brs, 1H), 8.56 (s, 1H), 8.24 (s, 1H), 8.14 (d, J=8.0 Hz, 1H), 7.38
(d, J=8.0 Hz, 1H), 7.25 (brd, 1H), 6.71 (brd, 1H), 4.64 (s, 2H),
3.93 (s, 3H).
Example 80
5-((9H-Purin-6-yl)amino)-4-methoxyisoindolin-1-one
##STR00113##
[0759] To the solution of 5-amino-4-methoxyisoindolin-1-one 80a (22
mg, 0.125 mmol) in acetic acid (2 mL) was added 6-chloro-9H-purine
(40 mg, 0.25 mmol) at room temperature. The reaction tube was
sealed and the resulting solution was stirred at 100.degree. C. for
1 hour. Then it was concentrated under reduced pressure.
Purification of the residue using Prep-HPLC (water (0.2% formic
acid), 10%.about.% acetonitrile, 15 minutes) gave
5-((9H-Purin-6-yl)amino)-4-methoxyisoindolin-1-one 80 (2.2 mg,
0.0074 mmol, white solid). Yield: 6%.
[0760] MS m/z (ESI): 297 [M+1],
[0761] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 13.62 (s, 1H), 9.71
(s, 1H), 8.66 (s, 1H), 8.31 (s, 1H), 8.12 (s, 1H), 7.87 (d, J=8.0
Hz, 1H), 7.40 (d, J=8.0 Hz, 1H), 4.62 (s, 2H), 3.90 (s, 3H).
Example 81
5-((1H-Pyrazolo[3,4-d]pyrimidin-4-yl)amino)-4-methoxyisoindolin-1-one
##STR00114##
[0763] The synthetic procedure is similar to that of example 74.
Using 4-chloro-1H-pyrazolo[3,4-d]pyrimidine instead of
6-chloro-9H-purine, the desired compound:
5-((1H-pyrazolo[3,4-d]pyrimidin-4-yl)amino)-4-methoxyisoindolin-1-one
(6 mg, 0.02 mmol, white solid) was obtained. Yield: 16%.
[0764] MS m/z (ESI): 297 [M+1],
[0765] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 13.32 (s, 1H),
8.68-8.84 (m, 2H), 8.57 (s, 1H), 8.46 (s, 1H), 8.34 (s, 1H), 7.42
(d, J=8.4 Hz, 1H), 4.65 (s, 2H), 4.05 (s, 3H).
Example 82
5-((5-Methylaminopyrimidin-4-yl)
amino)-4-methoxyisodihydroindole-1-one formate
##STR00115##
[0767] 4-Chloro-N-methylpyrimidin-5-amine 82a (10 mg, 0.070 mmol),
4-methoxy-5-((5-(methylamino) pyrimidin-4-yl) amino)
isodihydroindole-1-one (15 mg, 0.084 mmol), hydrochloric acid in
methanol (0.5 mL, 2 M) and butan-1-ol (0.5 mL) were mixed and
stirred at 130.degree. C. for 2 hours. Triethylamine was added
until pH=7.about.8. Then it was concentrated under reduced
pressure. Purification of the residue using Prep-HPLC (water (0.2%
formic acid), 20%.about.60% acetonitrile, 15 minutes) gave
5-((5-methylaminopyrimidin-4-yl)
amino)-4-methoxyisodihydroindole-1-one formate 82 (2.0 mg, 0.007
mmol, white solid).
[0768] Yield: 10%.
[0769] MS m/z (ESI): 286 [M+1],
[0770] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.26 (d, J=8.4 Hz,
1H), 8.17 (s, 1H), 7.73 (s, 1H), 7.51 (d, J=8.4 Hz, 1H), 4.68 (s,
2H), 4.01 (s, 3H), 2.91 (s, 3H).
Example 83
5-((6-((3-Chloropyridin-2-yl)amino)pyrimidin-4-yl)amino)-4-methoxyisoindol-
in-1-one
##STR00116##
[0772] The synthetic procedure is similar to that of example 78.
Using 6-chloro-N-(3-chloropyridin-2-yl)pyrimidine-4-amine instead
of 6-chloro-N-(pyridin-2-yl)pyrimidine-4-amine, the desired
compound
5-((6-((3-Chloropyridin-2-yl)amino)pyrimidin-4-yl)amino)-4-methoxyisoindo-
lin-1-one was obtained. Yield: 17%.
[0773] MS m/z (ESI): 383 & 385 [M+1],
[0774] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.01 (brs, 1H),
8.51 (brs, 1H), 8.32 (s, 1H), 8.31 (d, J=8.0 Hz, 1H), 8.14 (d,
J=8.0 Hz, 1H), 7.96 (d, J=8.0 Hz, 1H), 7.59 (s, 1H), 7.34 (d, J=8.0
Hz, 1H), 7.12-7.09 (m, 1H), 6.05 (brs, 1H), 4.56 (s, 2H), 3.91 (s,
3H).
Example 84
5-((6-(Pyrazin-2-ylamino) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindol-1-one formate
##STR00117##
[0776] The synthetic procedure is similar to that of example 69.
Using 2-chloropyrazine instead of 2-chloropyrimidine, the desired
compound: 5-((6-(pyrazin-2-ylamino) pyrimidin-4-yl)
amino)-4-methoxyisodihydroindol-1-one formate was obtained.
[0777] Yield: 4%.
[0778] MS m/z (ESI): 350 [M+1],
[0779] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.21 (s, 1H), 9.03
(s, 1H), 8.85 (s, 1H), 8.55 (s, 1H), 8.35 (s, 1H), 8.29 (s, 1H),
8.23 (s, 1H), 8.15 (s, 1H), 8.11 (d, J=8.2 Hz, 1H), 7.42 (s, 1H),
7.35 (d, J=8.2 Hz, 1H), 4.57 (s, 2H), 3.92 (s, 3H).
Example 85
5-((6-((5-Chloropyridin-2-yl)amino)pyrimidin-4-yl)amino)-4-methoxyisoindol-
in-1-one
##STR00118##
[0781] The synthetic procedure is similar to that of example 69.
Using 2, 5-dichloropyridine instead of 2-chloropyrimidine, the
desired compound:
5-((6-((5-Chloropyridin-2-yl)amino)pyrimidin-4-yl)amino)-4-methoxyisoindo-
lin-1-one was obtained. Yield: 9%.
[0782] MS m/z (ESI): 383 & 385 [M+1],
[0783] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.02 (s, 1H), 8.92
(s, 1H), 8.51 (s, 1H), 8.33 (s, 1H), 8.28 (s, 1H), 8.08 (d, J=8.0
Hz, 1H), 7.80 (d, J=8.8 Hz, 1H), 7.62 (d, J=8.8 Hz, 1H), 7.34 (d,
J=8.0 Hz, 1H), 7.32 (s, 1H), 4.56 (s, 2H), 3.91 (s, 3H).
Example 86
5-((6-((6-Chloropyridin-2-yl)amino)pyrimidin-4-yl)amino)-4-methoxyisoindol-
in-1-one
##STR00119##
[0785] The synthetic procedure is similar to that of example 69.
Using 2, 6-chloropyridine instead of 2-chloropyrimidine, the
desired compound:
5-((6-((6-Chloropyridin-2-yl)amino)pyrimidin-4-yl)amino)-4-methoxyisoindo-
lin-1-one was obtained. Yield: 9%.
[0786] MS m/z (ESI): 383 & 385 [M+1],
[0787] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.16 (s, 1H), 8.96
(s, 1H), 8.54 (s, 1H), 8.34 (s, 1H), 7.90 (d, J=7.2 Hz, 1H), 7.74
(t, J=8.0 Hz, 1H), 7.66 (d, J=8.0 Hz, 1H), 7.35 (d, J=8.0 Hz, 1H),
7.11 (s, 1H), 7.00 (d, J=7.2 Hz, 1H), 4.57 (s, 2H), 3.91 (s,
3H).
Example 87
5-((6-((4-Chloropyridin-2-yl)amino)pyrimidin-4-yl)amino)-4-methoxyisoindol-
in-1-one
##STR00120##
[0789] The synthetic procedure is similar to that of example 69.
Using 2-fluorine-4-chloropyridine instead of 2-chloropyrimidine,
the desired compound:
5-((6-((4-Chloropyridin-2-yl)amino)pyrimidin-4-yl)amino)-4-meth-
oxyisoindolin-1-one was obtained. Yield: 10%.
[0790] MS m/z (ESI): 383 & 385 [M+1],
[0791] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.99 (s, 1H), 8.89
(s, 1H), 8.45 (s, 1H), 8.29 (s, 1H), 8.17 (d, J=5.6 Hz, 1H), 8.00
(d, J=8.0 Hz, 1H), 7.69 (s, 1H), 7.27 (d, J=8.0 Hz, 1H), 7.26 (s,
1H), 6.98 (d, J=5.6 Hz, 1H), 4.49 (s, 2H), 3.84 (s, 3H).
[0792] MNK1 Inhibitory Activity Assay:
[0793] Evaluation of the effects of compounds of the invention on
mitogen-activated protein kinase kinase 1 (MNK1) activity using in
vitro kinase assays.
[0794] The experimental methods are summarized as follows: The in
vitro activity of MNK1 was determined by measuring the level of ADP
produced in the kinase reaction using the ADP-Glo Kinase Assay Kit.
The reaction buffer consisted of the following components: 50 mM
HEPES, pH 7.5, 10 mM MgCl.sub.2, 1 mM EGTA, 0.01% Brij35. Human
recombinant full-length MNK1 protein (Thermo, Cat. No. PR9138A) was
diluted with a reaction buffer to a 3.13 ng/uL kinase solution. The
substrate reaction solution consisted of a substrate diluted to
0.75 mg/ml in reaction buffer (GRSRSRSRSR, available from Scilight
Domestic Company), 2250 uM ATP, the ADP-Glo reagent and kinase
assay solution from Promega kit (Promega, V9102).
[0795] Compounds were diluted in 100% DMSO to 100 uM, then serially
diluted in 4 fold each time with DMSO to a minimum concentration of
0.0061 uM, and each concentration point was diluted 20-fold with
the reaction buffer. If the compound IC50 value is very low, the
initial concentration of the compound can be lowered. To a 384-well
assay plate (Thermo, Cat. No. 264706) were added 1 uL of compound
solution and 2 uL of MNK1 kinase solution. The mixture was mixed
well and incubated for 30 minutes at room temperature; then 2 uL of
substrate reaction solution was added, the reaction mixture was
incubated for 120 minutes at room temperature. The reaction was
terminated after an equal volume of 5uLADP-Glo solution was added,
and the remaining ATP was completely consumed. After mixing for 60
minutes at room temperature, 10 ul of the kinase assay solution was
added, and the mixture was uniformly mixed, and allowed to stand at
room temperature for 40 minutes in the dark. The detection solution
converts ADP into new ATP through coupled luciferase/luciferin, and
ATP is converted into optical signal by Ultr-Glo.TM. luciferase,
which can be detected by Envision. The intensity of the light
signal is proportional correlated with the amount of ADP produced
in the kinase reaction, therefore, the activity of MNK1 kinase can
be determined. In this experiment, the group without protein was
used as a negative control (100% inhibition). The group with
protein but without compounds was used as a positive control (0%
inhibition).
[0796] The percent inhibition of MNK1 activity by a compound can be
calculated by the following formula:
Inhibition
percentage=100-100*(signal.sub.compound-signal.sub.negative
control)/(signal.sub.positive control-signal.sub.negative
control
[0797] Compound IC.sub.50 value was calculated by the following
formula using 10 concentration points and XLfit (ID Business
Solutions Ltd., UK) software
Y=Bottom+(Top-Bottom)/(1+10{circumflex over ( )}((Log
IC.sub.50-X)*slope factor))
[0798] where Y is inhibition percentage. Bottom is the bottom
plateau of the curve, Top is the top plateau of the curve, and X is
the logarithm of the concentration of the test compound.
[0799] The results of the enzyme experiment test are shown in Table
1 below.
TABLE-US-00001 TABLE 1 Compound MNK1 IC50 Number (nM) 1 15.5 2 59.5
3 11.3 4 1.45 5 585.3 6 29.8 7 44.9 8 64.1 9 294.1 10 23.8 11
531.76 12 729.5 13 224.7 14 1105.9 15 21.0 16 n/a 17 27.9 18 10.5
19 45.0 20 29.2 21 78.2 22 23.3 23 31.3 24 42.2 25 40.0 26 36.6 27
43.8 28 539.2 29 57.9 30 140.7 31 126.3 32 165.6 33 94.7 34 450.3
35 32.5 36 7.8 37 23.8 38 11.9 39 13.0 40 1137.1 41 1048.4 42 600.1
43 264.5 44 1004.4 45 35.3 46 49.4 47 79.1 48 187.0 49 1052.0 50
253.9 51 197.5 52 14.4 53 22.8 54 34.3 55 187.4 56 19.8 57 323.1 58
6.1 59 432.2 60 792.3 61 593.8 62 75.3 63 97.1 64 437.2 65 693.8 66
38.5 67 2.7 68 8.2 69 27.3 70 17.0 71 128.1 72 30.5 73 188.3 74
38.4 75 34.4 76 571.9 77 899.7 78 19.7 79 20.9 80 11.7 81 102.7 82
154.2 83 96.6 84 5.0 85 13.9 86 18.5 87 19.1
MV-4-11 Cell Proliferation Inhibition Assay:
[0800] The effect of the compounds of the present invention on the
proliferation of MV-4-11 cells was evaluated using a luminescent
cell viability assay.
[0801] The experimental methods are summarized as follows:
[0802] The cell proliferation status of MV-4-11 was detected using
the CellTilter-Glo (CTG) assay kit: the luminescence signal
generated in the assay is proportional to the number of viable
cells in the culture medium by using a unique, stable luciferase to
detect viable cell metabolism indicator ATP.
[0803] CellTilter-Glo reagent (Promega, G7572) consists of
CellTilter-Glo lyophilized powder and ellTilter-Glo buffer, which
can be used to dissolve the lyophilized powder into the buffer.
[0804] MV-4-11 cells (ATCC #CRL-9591, purchased from Nanjing
Kezhen, item number CBP60522) were cultured in IMDM complete medium
(Thermofisher, 12440053) containing 10% FBS (GBICO, 10099-141) and
100 units/ml streptomycin mixture (Thermofisher, 15140122). when
the cells coverage rearched 80-90% in the culture container, dilute
and plant with 0.25% trypsin (including EDTA) (Thermofisher,
25200056) in a white 384-well plate (Thermofisher, 164610), 400
cells per well (36 .mu.l DMEM complete medium), then 384-well
plates were incubated overnight (18-20 hours) in a 37.degree. C., %
CO2 incubator.
[0805] Compounds were solubilized to 10 mM in 100% DMSO, then
sequentially diluted in 4-fold with DMSO to a minimum concentration
of 0.61 mM, and each concentration was diluted 50-fold with
FBS-free IMDM medium. If the compound IC50 value is very low, the
initial concentration of the compound can be lowered. 4 .mu.l of
the diluted IMDM compound was added to each well and mixed gently
by centrifugation. The GI50 (50% Growth Inhibition) of the
compounds was determined in this experiment, including the T0 group
before the cell proliferation (including the T0 positive and T0
negative control groups), and the T5 group after 120 hours of cell
proliferation (containing the T5 positive and T5 negative control
groups).). T0 represents the number of cells before cell
proliferation before compound addition, including positive control:
the group with the addition of cells and 0.2% DMSO was used as the
positive control. The group with medium alone was used as the
negative control, T0 group before the addition of compound will be
used for CTG test. In addition, the same positive and negative
groups were prepared and used as the control groups for cell
proliferation after 120 hours. The 384-well plate was placed in a
37.degree. C., 5% CO2 incubator for further incubation. After 120
hours, it was taken out and allowed to stand at room temperature
for 30 minutes. The CTG reagent was also taken out to room
temperature. 20 .mu.l of CTG reagent was added to each well, and
placed on a shaker. Gently shake for 5 minutes to ensure sufficient
cell lysis, leave for 10 minutes to stabilize the luminescence
signal, and then read the luminescence signal with EnVision (Perkin
Elmer).
[0806] The inhibition of MV-4-11 cell proliferation of the
compounds was calculated by the following formula:
Inhibition percentage=100-100*[(signal T5compound-signal
T5negative)-(signal T0positive-signal T0negative)]/[(signal
T5positive control-signal T5negative control)-(signal
T0positive-signal T0negative)]
[0807] The IC50 value of the compound was calculated from the 8
concentration points by XLfit (ID Business Solutions Ltd., UK)
software using the following formula:
Y=Bottom+(Top-Bottom)/(1+10{circumflex over ( )}((Log GI50-X)*slope
factor))
[0808] where Y is the percent inhibition and Bottom is the bottom
plateau of the curve, Top is the top plateau of the curve, and X is
the logarithm of the concentration of the test compound.
[0809] Cellular assays showed that the compounds of the examples of
the present invention have a significant inhibitory effect on the
proliferation of MV-4-11 tumor cells (IC.sub.50<500 nM), as
shown in Table 2 below
TABLE-US-00002 TABLE 2 Compound number MV-4-11 IC50 (nM) 4 301 67
483 68 295 84 278.
* * * * *